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transfection [2022/12/19 04:03]
pamela [Investigating the Function of DNA Sequences] 		transfection [2022/12/19 08:34] (current)
liam
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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.- Wikipedia 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.- Wikipedia
  
 +Transfection, technique used to insert foreign nucleic acid ( DNA or RNA) into a cell, typically with the intention of altering the properties of the cell. The introduction of nucleic acid from a different cell type can be accomplished using various biological, chemical, or physical methods. - Britannica 
 +{{ ::transfection_researchgate_2.png?400|  }}
 Transfection is the process of introducing foreign DNA into the cells either by physical (electroporation) or chemical (cationic lipid or calcium phosphate reagents) methods. Transfection is the process of introducing foreign DNA into the cells either by physical (electroporation) or chemical (cationic lipid or calcium phosphate reagents) methods.
 From: Stem Cell Manufacturing, 2016 From: Stem Cell Manufacturing, 2016
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 +Transfection is a modern and powerful method used to insert foreign nucleic acids into eukaryotic cells. The ability to modify host cells' genetic content enables the broad application of this process in studying normal cellular processes, disease molecular mechanism and gene therapeutic effect. [[:National Institutes of Health]] Apr 21, 2021 ((www.ncbi.nlm.nih.gov/pmc/articles/PMC8067914))
  
  
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 Cornel Mülhardt, E.W. Beese M.D., in Molecular Biology and Genomics, 2007 Cornel Mülhardt, E.W. Beese M.D., in Molecular Biology and Genomics, 2007
 Transfection through Bombardment: Particle Delivery Transfection through Bombardment: Particle Delivery
 +{{ ::transfection_researchgate_1.png?400|}}
  
 Transfection through **bombardment** was originally developed for the transfection of [[:plant cells]], but it is now also used for mammalian cells. Small wolfram or gold particles are coated with [[:DNA]] and then shot onto the cells at a high speed.  Transfection through **bombardment** was originally developed for the transfection of [[:plant cells]], but it is now also used for mammalian cells. Small wolfram or gold particles are coated with [[:DNA]] and then shot onto the cells at a high speed. 
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 +==== Graphene Based Gene Transfection ====
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 +Abstract
 +{{ ::graphine_based_gene_transfection_study_2010-_2.png?400|}}
 +Graphene as a star in materials research has been attracting tremendous attentions in the past few years in various fields including [[:biomedicine]]. In this work, for the first time we successfully use graphene as a [[:non-toxic nano-vehicle]] for efficient gene transfection. 
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 +[[:Graphene oxide]] (GO) is bound with cationic polymers, polyethyleneimine ([[:PEI]]) with two different molecular weights at 1.2 kDa and 10 kDa, forming GO-PEI-1.2k and GO-PEG-10k complexes, respectively, both of which are stable in physiological solutions. 
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 +Cellular toxicity tests reveal that our GO-PEI-10k complex exhibits significantly reduced toxicity to the treated cells compared to the bare PEI-10k polymer. The positively charged GO-PEI complexes are able to further bind with [[:plasmid DNA]] (pDNA) for intracellular transfection of the enhanced green fluorescence protein (EGFP) gene in [[:HeLa cells]]. 
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 +While EGFP transfection with PEI-1.2k appears to be ineffective, high EGFP expression is observed using the corresponding GO-PEI-1.2k as the transfection agent. On the other hand, GO-PEI-10k shows similar EGFP transfection efficiency but lower toxicity compared with PEI-10k. 
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 +Our results suggest graphene to be a [[:novel gene delivery nano-vector]] with low [[:cytotoxicity]] and high transfection efficiency, promising for future applications in non-viral based [[:gene therapy]]. ((https://web.archive.org/web/20220425062253/https://pubs.rsc.org/en/content/articlehtml/2011/nr/c0nr00680g?page=search))
 +
 +==== Jessica Rose Graphene Oxide Transfection ====
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 +Substack December 16,2022 by Jessica Rose
 +{{ ::graphine_transfection_jessica_rose_graphic.png?400|}}
 +Please refer to the published work in the journal ‘Nanoscale’ entitled: “Functionalized graphene oxide serves as a novel vaccine nano-adjuvant for robust stimulation of cellular immunity”. 
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 +This work was published in 2016 by a group in China and let me tell you, they did a lot of work and asked a lot of questions. Their methods are so good, that I could actually reproduce their work.
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 +Their primary goal was to find out if their adjuvant antigen delivery system, graphene oxide-polyethylene glycol-polyethylenimine ([[:GO-PEG-PE]]I), would be functional with regard to specific immune stimulation, and/or less toxic than aluminum-based adjuvants. 
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 +Their results showed that a PEG-PEI coated graphene oxide sheet - which incidentally really loves absorbing antigens like Ure B (an antigen associated with a bacterium called Helicobacter pylori) - was really good at trafficking antigen to dendritic cells to induce their maturation and activation when this concoction was injected intradermally. 
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 +What I really need to get my hands on are the original methods… this isn’t even locatable using sci-hub. Anyone crafty enough to get this paper? I have the supplementary materials but not the text and will not be providing personal details to ‘sign up’ for anything to read science. It’s against my code
  
 +Summary of paper translated into english: PEG-PEI-conjugated GO sheets used as adjuvants to deliver immune-stimulating agents like bacterial proteins, are a thing.
 +((https://jessicar.substack.com/p/graphene-oxide-conjugated-to-peg))
  
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