Stable Transfection

Transient and Stable Transfection

Capability to integrate genes into the full DNA sequence of a mammalian cell has a significant impact on biomedical research. Transfected genetic material can be expressed in the target cells either transiently or permanently depending on the methods utilized and the experimental questions being investigated.Transient transfections are used most commonly to analyze the short term impact of altered gene or protein expression. Plasmid DNA (pDNA), messenger RNA (mRNA), short interfering RNA (siRNA), and microRNA (miRNA), are introduced and gene products are expressed in the target cells however the nucleic acids do not integrate into the host cell genome. Therefore, gene product expression is transient and typically results in high expression levels that persist for 24-72 hours when RNA is transfected, or 48-96 hours following DNA transfection.

Conversely, in order to analyze the long term impact of altered gene or protein expression investigators typically utilize stable transfection protocols to develop stable cell lines. In a subpopulation of transfected cells, whether the desired effect is a stable or transient transfection, the transfected genetic material will integrate into the genome. In order to create stable cell lines, investigators will take advantage of this natural occurrence, and introduce the gene of interest along with a selectable marker. Therefore growth of transfected cells, in the presence of a selecting agent, will enable the subpopulation of cell where the exogenous genetic material has been incorporated into the genome to persist while the remaining cells undergo selection. Utilizing this method, investigators are able to develop cells that permanently express specific genes through their incorporation in the cellular genome.

Altogen Biosystems offer generation of stable cell lines service by transforming your cell line to stably express vector or gene of interest.

In Vivo Transfection Kits from Altogen Biosystems

RNAi has been used for in vivo target validation studies using animal models. The major challenge in performing RNAi studies in vivo is the effective, directed delivery of functional siRNA, shRNA, and miRNA molecules into specific tissues. ALTOGEN® In Vivo Transfection Reagents could be conjugated with siRNA and administered intratumorally (i.t) or systemically via intravenous (i.v) tail vein injection in order to provide directed gene silencing in specific tissues, including liver, pancreas, kidney, and tumors. Selective knockdown could be seen as early as 24 hours after injection.

Selected in vivo transfection product citations (ALTOGEN® IN VIVO Transfection Kits used in the following publications):

  • Nature. 2008 454(7203):523-7. Innate immunity induced by composition-dependent RIG-I ...Saito et al [PDF]
  • Am J Pathology. 2010 177(4):1870-80. Role of ocular complement factor H in a murine model ... Lyzogubov et al [PDF]
  • Nature Biotechnology. 2011 29(4):341-5. Delivery of siRNA to the mouse brain by ... Alvarez-Erviti et al [PDF]
  • Cancer Research. 2011 71(15):5144-53. Inhibition of miR-193a expression by... Iliopoulos et al [PDF]
  • RNA. 2010 16(11):2108-19. RNase L releases a small RNA from HCV RNA that refolds ... Malathi et al [PDF]
  • Diabetologia. 2012 55(7):2069-79. The p47phox- and NADPH oxidase organiser 1 ... Youn et al [PDF]
  • British Journal of Cancer. 2012 107(3):516-26. TIGAR induces p53-mediated cell-cycle ... Madan et al [PDF]
  • Hypertension. 2014 63(2):353-61. Tissue transglutaminase contributes to ... Liu et al [PDF]
  • Circulation Research. 2010 15;107(8). Kruppel-like factor-4 transcriptionally regulates ... Cowan et al [PDF]
  • Hypertension. 2012 59(1):158-66. Role of uncoupled endothelial nitric oxide synthase ... Gao et al [PDF]
  • Jounal of Biological Chemistry. 2012 287(4):2907. Chaperoning of mutant p53 protein ... Gogna et al [PDF]
  • PLoS Pathogens. 2012 8(8) Uridine composition of the poly-U/UC tract of HCV RNA ... Schnell et al [PDF]
  • J Proteome Res. 2012(11) Retinal proteome analysis in a mouse model of oxygen-induced ... Kim et al [PDF]
  • J Transl Med. 2010 15;8:133. Prevention of hyperglycemia-induced myocardial apoptosis ... Zhang et al [PDF]
  • Mol Cell Biol. 2013 33(7). SCO2 induces p53-mediated apoptosis by Thr845 phosphorylation ... Madan et al [PDF]
  • Hypertension. 2015 65(2):430-9. Neurokinin 3 receptor and phosphocholine transferase... Parchim et al [PDF]
  • Gastroenterology. 2011 141(2) Differential type I interferon-mediated autophagic trafficking ... Desai et al [PDF]
  • PLoS Pathog. 2014 10(10) Exosomes from hepatitis C infected patients transmit HCV ... Bukong et al [PDF]

Featured articles:

siRNA Transfection || Transfection Methods || Stable Transfection || RNAi || siRNA Library Screening || RNAi Therapeutics || In Vivo Transfection Reagents || Cell Line Specific Transfection Reagents || Cell Transfection

Transfection Services by Altogen Labs CRO:

Altogen Labs CRO  offers biotechnology and pharmaceutical contract research services, including: in vivo toxicology, xenograft services, generation of stable cell lines, RNA interference (RNAi) services, pharmacology and toxicology testing: IC-50, ELISA assay development, siRNA library screening and transfection services.

Featured transfection products from Altogen Biosystems:

In Vivo Transfection Kits || Fibroblast Cell Transfection Kit || HepG2 Cell Transfection Reagent || MDA-MB Transfection Reagent || A549 Cell Transfection Reagent || HUVEC Transfection Reagent || LNCaP Cell Transfection Kit ||  In Vivo Nanoparticle Transfection Kit  ||  In Vivo PEG-Liposome Transfection Kit  ||  In Vivo Lipid Transfection Kit  ||  In Vivo Polymer Transfection Kit 

Altogen Biosystems is a life sciences company dedicated to the development, marketing and manufacture of cell type specific transfection reagents. Efficient delivery of DNA, RNA, and siRNA enabled by advanced formulation of reagents and peculiar design of protocols. Altogen Biosystems offers a complete transfection system for a broad range of cell lines. All reagents are functionally tested to be highly reproducible, serum compatible, induce low toxicity, and can be used for co-transfection experiments, and high throughput applications.