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Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. It may also refer to other methods and cell types, although other terms are often preferred: "transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryotic cells, including plant cells. In animal cells, transfection is the preferred term as transformation is also used to refer to progression to a cancerous state (carcinogenesis) in these cells. Transduction is often used to describe virus-mediated gene transfer into eukaryotic cells.

The word transfection is a portmanteau of trans- and infection. Genetic material (such as supercoiled plasmid DNA or siRNA constructs), may be transfected. Transfection of animal cells typically involves opening transient pores or "holes" in the cell membrane to allow the uptake of material. Transfection can be carried out using calcium phosphate (i.e. tricalcium phosphate), by electroporation, by cell squeezing, or by mixing a cationic lipid with the material to produce liposomes that fuse with the cell membrane and deposit their cargo inside.

Transfection can result in unexpected morphologies and abnormalities in target cells.

Contents

The meaning of the term has evolved. The original meaning of transfection was "infection by transformation", i.e., introduction of genetic material, DNA or RNA, from a prokaryote-infecting virus or bacteriophage into cells, resulting in an infection. For work with bacterial and archaeal cells transfection retains its original meaning as a special case of transformation. Because the term transformation had another sense in animal cell biology (a genetic change allowing long-term propagation in culture, or acquisition of properties typical of cancer cells), the term transfection acquired, for animal cells, its present meaning of a change in cell properties caused by introduction of DNA.[citation needed]

There are various methods of introducing foreign DNA into a eukaryotic cell: some rely on physical treatment (electroporation, cell squeezing, nanoparticles, magnetofection); others rely on chemical materials or biological particles (viruses) that are used as carriers. There are many different methods of gene delivery developed for various types of cells and tissues, from bacterial to mammalian. Generally, the methods can be divided into three categories: physical, chemical, and biological.

Physical methods include electroporation, microinjection, gene gun, impalefection, hydrostatic pressure, continuous infusion, and sonication. Chemicals include methods such as lipofection, which is a lipid-mediated DNA-transfection process utilizing liposome vectors. It can also include the use of polymeric gene carriers (polyplexes). Biological transfection is typically mediated by viruses, utilizing the ability of a virus to inject its DNA inside a host cell. A gene that is intended for delivery is packaged into a replication-deficient viral particle. Viruses used to date include retrovirus, lentivirus, adenovirus, adeno-associated virus, and herpes simplex virus.[citation needed]

Physical methods

Electroporator with square wave and exponential decay waveforms for in vitro, in vivo, adherent cell and 96 well electroporation applications. Manufactured by BTX Harvard Apparatus, Holliston MA USA.

Physical methods are the conceptually simplest, using some physical means to force the transfected material into the target cell's nucleus. The most widely used physical method is electroporation, where short electrical pulses disrupt the cell membrane, allowing the transfected nucleic acids to enter the cell. Other physical methods use different means to poke holes in the cell membrane: Sonoporation uses high-intensity ultrasound (attributed mainly to the cavitation of gas bubbles interacting with nearby cell membranes), optical transfection uses a highly focused laser to form a ~1 µm diameter hole.

Several methods use tools that force the nucleic acid into the cell, namely: microinjection of nucleic acid with a fine needle; biolistic particle delivery, in which nucleic acid is attached to heavy metal particles (usually gold) and propelled into the cells at high speed; and magnetofection, where nucleic acids are attached to magnetic iron oxide particles and driven into the target cells by magnets.

Hydrodynamic delivery is a method used in mice and rats, in which nucleic acids can be delivered to the liver by injecting a relatively large volume in the blood in less than 10 seconds; nearly all of the DNA is expressed in the liver by this procedure.

Chemical methods

Chemical-based transfection can be divided into several kinds: cyclodextrin, polymers, liposomes, or nanoparticles (with or without chemical or viral functionalization. See below).

  • One of the cheapest methods uses calcium phosphate, originally discovered by F. L. Graham and A. J. van der Eb in 1973 (see also). HEPES-buffered saline solution (HeBS) containing phosphate ions is combined with a calcium chloride solution containing the DNA to be transfected. When the two are combined, a fine precipitate of the positively charged calcium and the negatively charged phosphate will form, binding the DNA to be transfected on its surface. The suspension of the precipitate is then added to the cells to be transfected (usually a cell culture grown in a monolayer). By a process not entirely understood, the cells take up some of the precipitate, and with it, the DNA. This process has been a preferred method of identifying many oncogenes.
  • Another method is the use of cationic polymers such as DEAE-dextran or polyethylenimine (PEI). The negatively charged DNA binds to the polycation and the complex is taken up by the cell via endocytosis.
  • Lipofection (or liposome transfection) is a technique used to inject genetic material into a cell by means of liposomes, which are vesicles that can easily merge with the cell membrane since they are both made of a phospholipid bilayer. Lipofection generally uses a positively charged (cationic) lipid (cationic liposomes or mixtures) to form an aggregate with the negatively charged (anionic) genetic material. This transfection technology performs the same tasks as other biochemical procedures utilizing polymers, DEAE-dextran, calcium phosphate, and electroporation. The efficiency of lipofection can be improved by treating transfected cells with a mild heat shock.
  • Fugene is a series of widely used proprietary non-liposomal transfection reagents capable of directly transfecting a wide variety of cells with high efficiency and low toxicity.
  • Dendrimer is a class of highly branched molecules based on various building blocks and synthesized through a convergent or a divergent method. These dendrimers bind the nucleic acids to form dendriplexes that then penetrate the cells.

Viral methods

DNA can also be introduced into cells using viruses as a carrier. In such cases, the technique is called transduction, and the cells are said to be transduced. Adenoviral vectors can be useful for viral transfection methods because they can transfer genes into a wide variety of human cells and have high transfer rates. Lentiviral vectors are also helpful due to their ability to transduce cells not currently undergoing mitosis.

Protoplast fusion is a technique in which transformed bacterial cells are treated with lysozyme in order to remove the cell wall. Following this, fusogenic agents (e.g., Sendai virus, PEG, electroporation) are used in order to fuse the protoplast carrying the gene of interest with the target recipient cell. A major disadvantage of this method is that bacterial components are non-specifically introduced into the target cell as well.

Stable and transient transfection differ in their long term effects on a cell; a stably-transfected cell will continuously express transfected DNA and pass it on to daughter cells, while a transiently-transfected cell will express transfected DNA for a short amount of time and not pass it on to daughter cells.

For some applications of transfection, it is sufficient if the transfected genetic material is only transiently expressed. Since the DNA introduced in the transfection process is usually not integrated into the nuclear genome, the foreign DNA will be diluted through mitosis or degraded. Cell lines expressing the Epstein–Barr virus (EBV) nuclear antigen 1 (EBNA1) or the SV40 large-T antigen, allow episomal amplification of plasmids containing the viral EBV (293E) or SV40 (293T) origins of replication, greatly reducing the rate of dilution.

If it is desired that the transfected gene actually remain in the genome of the cell and its daughter cells, a stable transfection must occur. To accomplish this, a marker gene is co-transfected, which gives the cell some selectable advantage, such as resistance towards a certain toxin. Some (very few) of the transfected cells will, by chance, have integrated the foreign genetic material into their genome. If the toxin is then added to the cell culture, only those few cells with the marker gene integrated into their genomes will be able to proliferate, while other cells will die. After applying this selective stress (selection pressure) for some time, only the cells with a stable transfection remain and can be cultivated further.

Common agents for selecting stable transfection are:

RNA can also be transfected into cells to transiently express its coded protein, or to study RNA decay kinetics. RNA transfection is often used in primary cells that do not divide.

siRNAs can also be transfected to achieve RNA silencing (i.e. loss of RNA and protein from the targeted gene). This has become a major application in research to achieve "knock-down" of proteins of interests (e.g. Endothelin-1) with potential applications in gene therapy. Limitation of the silencing approach are the toxicity of the transfection for cells and potential "off-target" effects on the expression of other genes/proteins.

RNA can be purified from cells after lysis or synthesized from free nucleotides either chemically, or enzymatically using an RNA polymerase to transcribe a DNA template. As with DNA, RNA can be delivered to cells by a variety of means including microinjection, electroporation, and lipid-mediated transfection. If the RNA encodes a protein, transfected cells may translate the RNA into the encoded protein. If the RNA is a regulatory RNA (such as a miRNA), the RNA may cause other changes in the cell (such as RNAi-mediated knockdown).

Encapsulating the RNA molecule in lipid nanoparticles was a breakthrough for producing viable RNA vaccines, solving a number of key technical barriers in delivering the RNA molecule into the human cell.

RNA molecules shorter than about 25nt (nucleotides) largely evade detection by the innate immune system, which is triggered by longer RNA molecules. Most cells of the body express proteins of the innate immune system, and upon exposure to exogenous long RNA molecules, these proteins initiate signaling cascades that result in inflammation. This inflammation hypersensitizes the exposed cell and nearby cells to subsequent exposure. As a result, while a cell can be repeatedly transfected with short RNA with few non-specific effects, repeatedly transfecting cells with even a small amount of long RNA can cause cell death unless measures are taken to suppress or evade the innate immune system (see "Long-RNA transfection" below).

Short-RNA transfection is routinely used in biological research to knock down the expression of a protein of interest (using siRNA) or to express or block the activity of a miRNA (using short RNA that acts independently of the cell's RNAi machinery, and therefore is not referred to as siRNA). While DNA-based vectors (viruses, plasmids) that encode a short RNA molecule can also be used, short-RNA transfection does not risk modification of the cell's DNA, a characteristic that has led to the development of short RNA as a new class of macromolecular drugs.

Long-RNA transfection is the process of deliberately introducing RNA molecules longer than about 25nt into living cells. A distinction is made between short- and long-RNA transfection because exogenous long RNA molecules elicit an innate immune response in cells that can cause a variety of nonspecific effects including translation block, cell-cycle arrest, and apoptosis.

Endogenous vs. exogenous long RNA

The innate immune system has evolved to protect against infection by detecting pathogen-associated molecular patterns (PAMPs), and triggering a complex set of responses collectively known as "inflammation". Many cells express specific pattern recognition receptors (PRRs) for exogenous RNA including toll-like receptor 3,7,8 (TLR3, TLR7, TLR8), the RNA helicase RIG1 (RARRES3), protein kinase R (PKR, a.k.a. EIF2AK2), members of the oligoadenylate synthetase family of proteins (OAS1, OAS2, OAS3), and others. All of these proteins can specifically bind to exogenous RNA molecules and trigger an immune response. The specific chemical, structural or other characteristics of long RNA molecules that are required for recognition by PRRs remain largely unknown despite intense study. At any given time, a typical mammalian cell may contain several hundred thousand mRNA and other, regulatory long RNA molecules. How cells distinguish exogenous long RNA from the large amount of endogenous long RNA is an important open question in cell biology. Several reports suggest that phosphorylation of the 5'-end of a long RNA molecule can influence its immunogenicity, and specifically that 5'-triphosphate RNA, which can be produced during viral infection, is more immunogenic than 5'-diphosphate RNA, 5'-monophosphate RNA or RNA containing no 5' phosphate. However, in vitro-transcribed (ivT) long RNA containing a 7-methylguanosine cap (present in eukaryotic mRNA) is also highly immunogenic despite having no 5' phosphate, suggesting that characteristics other than 5'-phosphorylation can influence the immunogenicity of an RNA molecule.

Eukaryotic mRNA contains chemically modified nucleotides such as N6-methyladenosine, 5-methylcytidine, and 2'-O-methylated nucleotides. Although only a very small number of these modified nucleotides are present in a typical mRNA molecule, they may help prevent mRNA from activating the innate immune system by disrupting secondary structure that would resemble double-stranded RNA (dsRNA), a type of RNA thought to be present in cells only during viral infection. The immunogenicity of long RNA has been used to study both innate and adaptive immunity.

Repeated long-RNA transfection

Inhibiting only three proteins, interferon-β, STAT2, and EIF2AK2 is sufficient to rescue human fibroblasts from the cell death caused by frequent transfection with long, protein-encoding RNA. Inhibiting interferon signaling disrupts the positive-feedback loop that normally hypersensitizes cells exposed to exogenous long RNA. Researchers have recently used this technique to express reprogramming proteins in primary human fibroblasts.

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Scholia has a profile for transfection (Q1429031).

Transfection Article Talk Language Watch Edit Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells 1 2 It may also refer to other methods and cell types although other terms are often preferred transformation is typically used to describe non viral DNA transfer in bacteria and non animal eukaryotic cells including plant cells In animal cells transfection is the preferred term as transformation is also used to refer to progression to a cancerous state carcinogenesis in these cells Transduction is often used to describe virus mediated gene transfer into eukaryotic cells 2 3 The word transfection is a portmanteau of trans and infection Genetic material such as supercoiled plasmid DNA or siRNA constructs may be transfected Transfection of animal cells typically involves opening transient pores or holes in the cell membrane to allow the uptake of material Transfection can be carried out using calcium phosphate i e tricalcium phosphate by electroporation by cell squeezing or by mixing a cationic lipid with the material to produce liposomes that fuse with the cell membrane and deposit their cargo inside Transfection can result in unexpected morphologies and abnormalities in target cells Contents 1 Terminology 2 Methods 2 1 Physical methods 2 2 Chemical methods 2 3 Viral methods 3 Stable and transient transfection 4 RNA transfection 4 1 Endogenous vs exogenous long RNA 4 2 Repeated long RNA transfection 5 See also 6 References 7 Further reading 8 External linksTerminology EditThe meaning of the term has evolved 4 The original meaning of transfection was infection by transformation i e introduction of genetic material DNA or RNA from a prokaryote infecting virus or bacteriophage into cells resulting in an infection For work with bacterial and archaeal cells transfection retains its original meaning as a special case of transformation Because the term transformation had another sense in animal cell biology a genetic change allowing long term propagation in culture or acquisition of properties typical of cancer cells the term transfection acquired for animal cells its present meaning of a change in cell properties caused by introduction of DNA citation needed Methods EditThere are various methods of introducing foreign DNA into a eukaryotic cell some rely on physical treatment electroporation cell squeezing nanoparticles magnetofection others rely on chemical materials or biological particles viruses that are used as carriers There are many different methods of gene delivery developed for various types of cells and tissues from bacterial to mammalian Generally the methods can be divided into three categories physical chemical and biological 5 Physical methods include electroporation microinjection gene gun impalefection hydrostatic pressure continuous infusion and sonication Chemicals include methods such as lipofection which is a lipid mediated DNA transfection process utilizing liposome vectors It can also include the use of polymeric gene carriers polyplexes 6 Biological transfection is typically mediated by viruses utilizing the ability of a virus to inject its DNA inside a host cell A gene that is intended for delivery is packaged into a replication deficient viral particle Viruses used to date include retrovirus lentivirus adenovirus adeno associated virus and herpes simplex virus citation needed Physical methods Edit Electroporator with square wave and exponential decay waveforms for in vitro in vivo adherent cell and 96 well electroporation applications Manufactured by BTX Harvard Apparatus Holliston MA USA Physical methods are the conceptually simplest using some physical means to force the transfected material into the target cell s nucleus The most widely used physical method is electroporation where short electrical pulses disrupt the cell membrane allowing the transfected nucleic acids to enter the cell 5 Other physical methods use different means to poke holes in the cell membrane Sonoporation uses high intensity ultrasound attributed mainly to the cavitation of gas bubbles interacting with nearby cell membranes optical transfection uses a highly focused laser to form a 1 µm diameter hole 7 Several methods use tools that force the nucleic acid into the cell namely microinjection of nucleic acid with a fine needle 5 biolistic particle delivery in which nucleic acid is attached to heavy metal particles usually gold and propelled into the cells at high speed 8 and magnetofection where nucleic acids are attached to magnetic iron oxide particles and driven into the target cells by magnets 8 Hydrodynamic delivery is a method used in mice and rats in which nucleic acids can be delivered to the liver by injecting a relatively large volume in the blood in less than 10 seconds nearly all of the DNA is expressed in the liver by this procedure 9 Chemical methods Edit Chemical based transfection can be divided into several kinds cyclodextrin 10 polymers 11 liposomes or nanoparticles 12 with or without chemical or viral functionalization See below One of the cheapest methods uses calcium phosphate originally discovered by F L Graham and A J van der Eb in 1973 13 see also 14 HEPES buffered saline solution HeBS containing phosphate ions is combined with a calcium chloride solution containing the DNA to be transfected When the two are combined a fine precipitate of the positively charged calcium and the negatively charged phosphate will form binding the DNA to be transfected on its surface The suspension of the precipitate is then added to the cells to be transfected usually a cell culture grown in a monolayer By a process not entirely understood the cells take up some of the precipitate and with it the DNA This process has been a preferred method of identifying many oncogenes 15 Another method is the use of cationic polymers such as DEAE dextran or polyethylenimine PEI The negatively charged DNA binds to the polycation and the complex is taken up by the cell via endocytosis Lipofection or liposome transfection is a technique used to inject genetic material into a cell by means of liposomes which are vesicles that can easily merge with the cell membrane since they are both made of a phospholipid bilayer 16 Lipofection generally uses a positively charged cationic lipid cationic liposomes or mixtures to form an aggregate with the negatively charged anionic genetic material 17 This transfection technology performs the same tasks as other biochemical procedures utilizing polymers DEAE dextran calcium phosphate and electroporation The efficiency of lipofection can be improved by treating transfected cells with a mild heat shock 18 Fugene is a series of widely used proprietary non liposomal transfection reagents capable of directly transfecting a wide variety of cells with high efficiency and low toxicity 19 20 21 22 Dendrimer is a class of highly branched molecules based on various building blocks and synthesized through a convergent or a divergent method These dendrimers bind the nucleic acids to form dendriplexes that then penetrate the cells 23 24 Viral methods Edit DNA can also be introduced into cells using viruses as a carrier In such cases the technique is called transduction and the cells are said to be transduced Adenoviral vectors can be useful for viral transfection methods because they can transfer genes into a wide variety of human cells and have high transfer rates 2 Lentiviral vectors are also helpful due to their ability to transduce cells not currently undergoing mitosis Protoplast fusion is a technique in which transformed bacterial cells are treated with lysozyme in order to remove the cell wall Following this fusogenic agents e g Sendai virus PEG electroporation are used in order to fuse the protoplast carrying the gene of interest with the target recipient cell A major disadvantage of this method is that bacterial components are non specifically introduced into the target cell as well Stable and transient transfection EditStable and transient transfection differ in their long term effects on a cell a stably transfected cell will continuously express transfected DNA and pass it on to daughter cells while a transiently transfected cell will express transfected DNA for a short amount of time and not pass it on to daughter cells For some applications of transfection it is sufficient if the transfected genetic material is only transiently expressed Since the DNA introduced in the transfection process is usually not integrated into the nuclear genome the foreign DNA will be diluted through mitosis or degraded 5 Cell lines expressing the Epstein Barr virus EBV nuclear antigen 1 EBNA1 or the SV40 large T antigen allow episomal amplification of plasmids containing the viral EBV 293E or SV40 293T origins of replication greatly reducing the rate of dilution 25 If it is desired that the transfected gene actually remain in the genome of the cell and its daughter cells a stable transfection must occur To accomplish this a marker gene is co transfected which gives the cell some selectable advantage such as resistance towards a certain toxin Some very few of the transfected cells will by chance have integrated the foreign genetic material into their genome If the toxin is then added to the cell culture only those few cells with the marker gene integrated into their genomes will be able to proliferate while other cells will die After applying this selective stress selection pressure for some time only the cells with a stable transfection remain and can be cultivated further 26 Common agents for selecting stable transfection are Geneticin or G418 neutralized by the product of the neomycin resistance gene Puromycin Zeocin Hygromycin B Blasticidin SRNA transfection EditRNA can also be transfected into cells to transiently express its coded protein or to study RNA decay kinetics RNA transfection is often used in primary cells that do not divide siRNAs can also be transfected to achieve RNA silencing i e loss of RNA and protein from the targeted gene This has become a major application in research to achieve knock down of proteins of interests e g Endothelin 1 27 with potential applications in gene therapy Limitation of the silencing approach are the toxicity of the transfection for cells and potential off target effects on the expression of other genes proteins RNA can be purified from cells after lysis or synthesized from free nucleotides either chemically or enzymatically using an RNA polymerase to transcribe a DNA template As with DNA RNA can be delivered to cells by a variety of means including microinjection electroporation and lipid mediated transfection If the RNA encodes a protein transfected cells may translate the RNA into the encoded protein 28 If the RNA is a regulatory RNA such as a miRNA the RNA may cause other changes in the cell such as RNAi mediated knockdown Encapsulating the RNA molecule in lipid nanoparticles was a breakthrough for producing viable RNA vaccines solving a number of key technical barriers in delivering the RNA molecule into the human cell 29 30 RNA molecules shorter than about 25nt nucleotides largely evade detection by the innate immune system which is triggered by longer RNA molecules Most cells of the body express proteins of the innate immune system and upon exposure to exogenous long RNA molecules these proteins initiate signaling cascades that result in inflammation This inflammation hypersensitizes the exposed cell and nearby cells to subsequent exposure As a result while a cell can be repeatedly transfected with short RNA with few non specific effects repeatedly transfecting cells with even a small amount of long RNA can cause cell death unless measures are taken to suppress or evade the innate immune system see Long RNA transfection below Short RNA transfection is routinely used in biological research to knock down the expression of a protein of interest using siRNA or to express or block the activity of a miRNA using short RNA that acts independently of the cell s RNAi machinery and therefore is not referred to as siRNA While DNA based vectors viruses plasmids that encode a short RNA molecule can also be used short RNA transfection does not risk modification of the cell s DNA a characteristic that has led to the development of short RNA as a new class of macromolecular drugs 31 Long RNA transfection is the process of deliberately introducing RNA molecules longer than about 25nt into living cells A distinction is made between short and long RNA transfection because exogenous long RNA molecules elicit an innate immune response in cells that can cause a variety of nonspecific effects including translation block cell cycle arrest and apoptosis Endogenous vs exogenous long RNA Edit The innate immune system has evolved to protect against infection by detecting pathogen associated molecular patterns PAMPs and triggering a complex set of responses collectively known as inflammation Many cells express specific pattern recognition receptors PRRs for exogenous RNA including toll like receptor 3 7 8 TLR3 TLR7 TLR8 32 33 34 35 the RNA helicase RIG1 RARRES3 36 protein kinase R PKR a k a EIF2AK2 37 38 members of the oligoadenylate synthetase family of proteins OAS1 OAS2 OAS3 and others All of these proteins can specifically bind to exogenous RNA molecules and trigger an immune response The specific chemical structural or other characteristics of long RNA molecules that are required for recognition by PRRs remain largely unknown despite intense study At any given time a typical mammalian cell may contain several hundred thousand mRNA and other regulatory long RNA molecules How cells distinguish exogenous long RNA from the large amount of endogenous long RNA is an important open question in cell biology Several reports suggest that phosphorylation of the 5 end of a long RNA molecule can influence its immunogenicity and specifically that 5 triphosphate RNA which can be produced during viral infection is more immunogenic than 5 diphosphate RNA 5 monophosphate RNA or RNA containing no 5 phosphate 39 40 41 42 43 44 However in vitro transcribed ivT long RNA containing a 7 methylguanosine cap present in eukaryotic mRNA is also highly immunogenic despite having no 5 phosphate 45 suggesting that characteristics other than 5 phosphorylation can influence the immunogenicity of an RNA molecule Eukaryotic mRNA contains chemically modified nucleotides such as N6 methyladenosine 5 methylcytidine and 2 O methylated nucleotides Although only a very small number of these modified nucleotides are present in a typical mRNA molecule they may help prevent mRNA from activating the innate immune system by disrupting secondary structure that would resemble double stranded RNA dsRNA 46 34 a type of RNA thought to be present in cells only during viral infection The immunogenicity of long RNA has been used to study both innate and adaptive immunity Repeated long RNA transfection Edit Inhibiting only three proteins interferon b STAT2 and EIF2AK2 is sufficient to rescue human fibroblasts from the cell death caused by frequent transfection with long protein encoding RNA 45 Inhibiting interferon signaling disrupts the positive feedback loop that normally hypersensitizes cells exposed to exogenous long RNA Researchers have recently used this technique to express reprogramming proteins in primary human fibroblasts 47 See also EditGene targeting Minicircle Protofection Transformation Transduction Transgene Vector molecular biology Viral vectorReferences Edit Transfection at the US National Library of Medicine Medical Subject Headings MeSH a b c Transfection Protocols and Applications Guide Promega Transduction Genetic at the US National Library of Medicine Medical Subject Headings MeSH Transfection at Dorland s Medical Dictionary a b c d Kim TK Eberwine JH August 2010 Mammalian cell transfection the present and the future Analytical and Bioanalytical Chemistry 397 8 3173 8 doi 10 1007 s00216 010 3821 6 PMC 2911531 PMID 20549496 Saul JM Linnes MP Ratner BD Giachelli CM Pun SH November 2007 Delivery of non viral gene carriers from sphere templated fibrin scaffolds for sustained transgene expression Biomaterials 28 31 4705 16 doi 10 1016 j biomaterials 2007 07 026 PMID 17675152 Tsukakoshi M Kurata S Nomiya Y et al 1984 A Novel Method of DNA Transfection by Laser Microbeam Cell Surgery Applied Physics B Photophysics and Laser Chemistry 35 3 135 140 Bibcode 1984ApPhB 35 135T doi 10 1007 BF00697702 S2CID 123250337 a b Mehier Humbert S Guy RH April 2005 Physical methods for gene transfer improving the kinetics of gene delivery into cells Adv Drug Deliv Rev 57 5 733 53 doi 10 1016 j addr 2004 12 007 PMID 15757758 Suda T Liu D 2015 Hydrodynamic delivery Adv Genet Advances in Genetics 89 89 111 doi 10 1016 bs adgen 2014 10 002 ISBN 9780128022726 PMID 25620009 Menuel S Fontanay S Clarot I Duval RE Diez L Marsura A December 2008 Synthesis and complexation ability of a novel bis guanidinium tetrakis beta cyclodextrin dendrimeric tetrapod as a potential gene delivery DNA and siRNA system Study of cellular siRNA transfection Bioconjugate Chemistry 19 12 2357 62 doi 10 1021 bc800193p PMID 19053312 Fischer D von Harpe A Kunath K Petersen H Li Y Kissel T 2002 Copolymers of ethylene imine and N 2 hydroxyethyl ethylene imine as tools to study effects of polymer structure on physicochemical and biological properties of DNA complexes Bioconjugate Chemistry 13 5 1124 33 doi 10 1021 bc025550w PMID 12236795 Nanoparticle Based Transfection Reagents Biology Transfection Research Resource Transfection ws Graham FL van der Eb AJ April 1973 A new technique for the assay of infectivity of human adenovirus 5 DNA Virology 52 2 456 67 doi 10 1016 0042 6822 73 90341 3 PMID 4705382 Bacchetti S Graham FL April 1977 Transfer of the gene for thymidine kinase to thymidine kinase deficient human cells by purified herpes simplex viral DNA Proceedings of the National Academy of Sciences of the United States of America 74 4 1590 4 Bibcode 1977PNAS 74 1590B doi 10 1073 pnas 74 4 1590 PMC 430836 PMID 193108 Kriegler M 1991 Transfer and Expression A Laboratory Manual W H Freeman pp 96 97 ISBN 978 0 7167 7004 6 Felgner PL Gadek TR Holm M Roman R Chan HW Wenz M Northrop JP Ringold GM Danielsen M November 1987 Lipofection a highly efficient lipid mediated DNA transfection procedure Proceedings of the National Academy of Sciences of the United States of America 84 21 7413 7 Bibcode 1987PNAS 84 7413F doi 10 1073 pnas 84 21 7413 PMC 299306 PMID 2823261 Felgner JH Kumar R Sridhar CN Wheeler CJ Tsai YJ Border R Ramsey P Martin M Felgner PL January 1994 Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations The Journal of Biological Chemistry 269 4 2550 61 doi 10 1016 S0021 9258 17 41980 6 PMID 8300583 Pipes BL Vasanwala FH Tsang TC Zhang T Luo P Harris DT January 2005 Brief heat shock increases stable integration of lipid mediated DNA transfections BioTechniques 38 1 48 52 doi 10 2144 05381bm05 PMID 15679084 Jacobsen LB Calvin SA Colvin KE Wright M June 2004 FuGENE 6 Transfection Reagent the gentle power Methods Transfection of Mammalian Cells 33 2 104 12 doi 10 1016 j ymeth 2003 11 002 PMID 15121164 Hellgren I Drvota V Pieper R Enoksson S Blomberg P Islam KB Sylven C August 2000 Highly efficient cell mediated gene transfer using non viral vectors and FuGene6 in vitro and in vivo studies Cellular and Molecular Life Sciences 57 8 9 1326 33 doi 10 1007 PL00000769 PMID 11028922 S2CID 27916034 Lakshmipathy U Thyagarajan B 2011 Primary and Stem Cells Gene Transfer Technologies and Applications 1st ed Wiley Blackwell ISBN 978 0 470 61074 9 Arnold AS Laporte V Dumont S Appert Collin A Erbacher P Coupin G Levy R Poindron P Gies JP February 2006 Comparing reagents for efficient transfection of human primary myoblasts FuGENE 6 Effectene and ExGen 500 Fundamental amp Clinical Pharmacology 20 1 81 9 doi 10 1111 j 1472 8206 2005 00344 x PMID 16448398 S2CID 42585711 Sapra Rachit Verma Ram P Maurya Govind P Dhawan Sameer Babu Jisha Haridas V 13 November 2019 Designer Peptide and Protein Dendrimers A Cross Sectional Analysis Chemical Reviews 119 21 11391 11441 doi 10 1021 acs chemrev 9b00153 ISSN 0009 2665 PMID 31556597 S2CID 203435702 Heitz Marc Javor Sacha Darbre Tamis Reymond Jean Louis 21 August 2019 Stereoselective pH Responsive Peptide Dendrimers for siRNA Transfection Bioconjugate Chemistry 30 8 2165 2182 doi 10 1021 acs bioconjchem 9b00403 ISSN 1043 1802 PMID 31398014 S2CID 199519310 Durocher Y Perret S Kamen A January 2002 High level and high throughput recombinant protein production by transient transfection of suspension growing human 293 EBNA1 cells Nucleic Acids Research 30 2 9e 9 doi 10 1093 nar 30 2 e9 PMC 99848 PMID 11788735 Fanelli A 2016 The Science of Stable Cell Line Generation Retrieved 23 December 2017 Mawji IA Marsden PA June 2006 RNA transfection is a versatile tool to investigate endothelin 1 posttranscriptional regulation Experimental Biology and Medicine 231 6 704 708 doi 10 3181 00379727 231 2310704 inactive 28 February 2022 PMID 16740984 a href wiki Template Cite journal title Template Cite journal cite journal a CS1 maint DOI inactive as of February 2022 link Herb M Farid A Gluschko A Kronke M Schramm M November 2019 Highly Efficient Transfection of Primary Macrophages with In Vitro Transcribed mRNA Journal of Visualized Experiments 153 doi 10 3791 60143 PMID 31762462 Cooney Elizabeth 1 December 2020 How nanotechnology helps mRNA Covid 19 vaccines work Stat Retrieved 3 December 2020 Foley Katherine Ellen 22 December 2020 The first Covid 19 vaccines have changed biotech forever Quartz Quartz Media Retrieved 11 January 2021 Tansey B 11 August 2006 Macular degeneration treatment interferes with RNA messages San Francisco Chronicle Alexopoulou L Holt AC Medzhitov R Flavell RA 2001 Recognition of double stranded RNA and activation of NF kappaB by Toll like receptor 3 Nature 413 6857 732 738 Bibcode 2001Natur 413 732A doi 10 1038 35099560 PMID 11607032 S2CID 4346537 Kariko K Ni H Capodici J Lamphier M Weissman D 2004 mRNA is an endogenous ligand for Toll like receptor 3 J Biol Chem 279 13 12542 12550 doi 10 1074 jbc M310175200 PMID 14729660 a b Kariko K Buckstein M Ni H Weissman D 2005 Suppression of RNA recognition by Toll like receptors the impact of nucleoside modification and the evolutionary origin of RNA Immunity 23 2 165 175 doi 10 1016 j immuni 2005 06 008 PMID 16111635 Diebold SS Kaisho T Hemmi H Akira S Reis e Sousa C 2004 Innate antiviral responses by means of TLR7 mediated recognition of single stranded RNA Science 303 5663 1529 1531 Bibcode 2004Sci 303 1529D doi 10 1126 science 1093616 PMID 14976261 S2CID 33144196 Yoneyama M Kikuchi M Natsukawa T Shinobu N Imaizumi T et al 2004 The RNA helicase RIG I has an essential function in double stranded RNA induced innate antiviral responses Nat Immunol 5 7 730 737 doi 10 1038 ni1087 PMID 15208624 S2CID 34876422 Das HK Das A Ghosh Dastidar P Ralston RO Yaghmai B et al 1981 Protein synthesis in rabbit reticulocytes Purification and characterization of a double stranded RNA dependent protein synthesis inhibitor from reticulocyte lysates J Biol Chem 256 12 6491 6495 doi 10 1016 S0021 9258 19 69192 1 PMID 7240221 Levin DH Petryshyn R London IM 1981 Characterization of purified double stranded RNA activated eIF 2 alpha kinase from rabbit reticulocytes J Biol Chem 256 14 7638 7641 doi 10 1016 S0021 9258 19 69008 3 PMID 6265457 Hornung V Ellegast J Kim S Brzozka K Jung A et al 2006 5 triphosphate RNA is the ligand for RIG I Science 314 5801 994 997 Bibcode 2006Sci 314 964H doi 10 1126 science 1132505 PMID 17038590 S2CID 22436759 Saito T Owen DM Jiang F Marcotrigiano J Gale M Jr 2008 Innate immunity induced by composition dependent RIG I recognition of Hepatitis C virus RNA Nature 454 7203 523 527 Bibcode 2008Natur 454 523S doi 10 1038 nature07106 PMC 2856441 PMID 18548002 Takahasi K Yoneyama M Nishihori T Hirai R Kumeta H et al 2008 Nonself RNA sensing mechanism of RIG I helicase and activation of antiviral immune responses Mol Cell 29 4 428 440 doi 10 1016 j molcel 2007 11 028 PMID 18242112 Yoneyama M Fujita T 2008 Structural mechanism of RNA recognition by the RIG I like receptors Immunity 29 2 178 181 doi 10 1016 j immuni 2008 07 009 PMID 18701081 Schmidt A Schwerd T Hamm W Hellmuth JC Cui S et al 2009 5 triphosphate RNA requires base paired structures to activate antiviral signaling via RIG I Proc Natl Acad Sci USA 106 29 12067 12072 Bibcode 2009PNAS 10612067S doi 10 1073 pnas 0900971106 PMC 2705279 PMID 19574455 Schlee M Roth A Hornung V Hagmann CA Wimmenauer V et al 2009 Recognition of 5 triphosphate by RIG I helicase requires short blunt double stranded RNA as contained in panhandle of negative strand virus Immunity 31 1 25 34 doi 10 1016 j immuni 2009 05 008 PMC 2824854 PMID 19576794 a b Angel M Yanik MF 2010 Innate Immune Suppression Enables Frequent Transfection with RNA Encoding Reprogramming Proteins PLOS ONE 5 7 e11756 Bibcode 2010PLoSO 511756A doi 10 1371 journal pone 0011756 PMC 2909252 PMID 20668695 Herb M Farid A Gluschko A Kronke M Schramm M November 2019 Highly Efficient Transfection of Primary Macrophages with In Vitro Transcribed mRNA Journal of Visualized Experiments 153 doi 10 3791 60143 PMID 31762462 Trafton A 26 July 2010 RNA offers a safer way to reprogram cells MIT News Office Further reading EditSegura T Shea LD 2001 Materials for non viral gene delivery Annual Review of Materials Research 31 25 46 Bibcode 2001AnRMS 31 25S doi 10 1146 annurev matsci 31 1 25 Luo D Saltzman WM January 2000 Synthetic DNA delivery systems Nature Biotechnology 18 1 33 7 doi 10 1038 71889 PMID 10625387 S2CID 7068508 Bonetta L 2005 The inside scoop evaluating gene delivery methods Nature Methods 2 11 875 883 doi 10 1038 nmeth1105 875 S2CID 8078059 External links EditScholia has a profile for transfection Q1429031 Transfection at the US National Library of Medicine Medical Subject Headings MeSH Biology Research Resource Articles and Forums about Transfection Research in optical transfection at the University of St Andrews The 10th US Japan Symposium on Drug Delivery Systems Retrieved from https en wikipedia org w index php title Transfection amp oldid 1085263014, wikipedia, wiki, book,

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