TransIT® Transfection Reagents for DNA and RNA Mediated Virus Production

DNA Mediated Virus Production Reagents

  • TransIT®-VirusGEN™: developed to provide high titer AAV & Lentivirus production with one reagent
  • TransIT®-Lenti: optimized for adherent Lentivirus production in 293T cells
  • TransIT®-LT1: the first serum compatible, broad spectrum transfection reagent
  • TransIT®-293: the first cell line specific reagent targeting efficient transfection of HEK-293 cells
  • TransIT®-2020: high efficiency broad spectrum, animal origin free transfection reagent

RNA Mediated Virus Production Reagents

  • TransIT®-mRNA: generate high titers with superior RNA virus delivery
 

DNA Mediated Virus Production

High Titer Virus Production Using TransIT-2020

Rescue of Influenza Virus by Reverse Transfection With TransIT®-2020 Transfection Reagent. 293T and MDBK cells are reverse transfected with up to 10 separate influenza virus rescue plasmids per experiment. Reagent:total DNA complexes are prepared with TransIT®-2020 Transfection Reagent at a 3:1 ratio of reagent to total DNA. Virus-containing supernatants are harvested 48-96 hr post-transfection. Titers from transfected cells can be as high as 107 pfu/ml. Rescued virus is then amplified on MDBK cells to increase titer and total yield. Viral stocks are subsequently titered by plaque assay on MDCK cells. Titers from representative rescue experiments with different viral strains and reassortants are shown. This experimental strategy is adapted from Neumann, et al. 2005 PNAS.

Data courtesy of Andrew Mehle, University of Wisconsin-Madison

TransIT-293 Reagent Achieves High Transfection Efficiency in HEK 293 Cells

TransIT®-293 Reagent Achieves High Transfection Efficiency in HEK 293 Cells. HEK 293 cells were transfected with pEGFP using TransIT®-293 Reagent in complete media. Cells were visualized under fluorescent microscopy 24 hours post-transfection.

 

Citations for DNA mediated virus production using TransIT® Transfection Reagents

Virus Cell Type Product Citation
AAV
293-F TransIT®-VirusGEN™ New! 
293T TransIT®-VirusGEN™ New! 
Lentivirus 293-F TransIT®-VirusGEN™ New! 
293T TransIT®-Lenti New!
Retrovirus 293T TransIT®-LT1 Moon et al. Hum Mol Genet. 2014;23(2):449-66.
Phoenix Eco TransIT®-LT1 Saxena et al./em>. Mol Cell Bio 2009; 29: 2335-2345
293FT TransIT®-293 Haxhinasto et al./em>. J Exp Med 2008; 205: 565-574.
Reovirus BHK-T7 TransIT®-LT1 Stebbing et al./em>. J. Virol. 2014 vol. 88: 2572-2583.
BHK-T7 TransIT®-LT1 Doyle et al./em>. J Biol Chem 2012; 287: 8029-8038.
L929 TransIT®-LT1 Danthi et al./em>. J Virol 2008; 82: 161-172.
Influenza virus 293T/MDBK TransIT®-2020 Tran et al. J. Virol. 2013; 87: 13321-13329.
293T TransIT®-LT1 Imai et al./em>. PLoS Path 2010; 6: e1001106.
293T/MDCK TransIT®-LT1 Xing et al./em>. J Gen Viol 2009; 90: 1109-1118.
Chandipura virus BSRT-7/5 TransIT®-LT1 Stock et al. J Gen Virol 2014; 95: 38-43.
HPV Pseudovirus 293TT TransIT®-2020 Jagu et al. J. Virol. 2013; 87: 6127-6136.
LACV and JCV/LACV BHK TransIT®-LT1 Bennett et al. J Virol 2012; 86: 420-426.
Epstein-Barr 293 TransIT®-293 Hughes et al./em>. J Virol 2012; 86: 1034-1045.
HIV 293T/C-33A TransIT®-LT1 L’hernault et al./em>. J Virol 2012; 86: 5867-5876.
293T TransIT®-LT1 Galli et al./em>. J Virol 2010; 84: 7651-7661.
Linearised Polyomavirus MasPyV genome BHK and Vero TransIT®-2020 Orba et al. J Gen Virol 2011; 92 no. 4 789-795.
VLP 293T TransIT®-293 Crist et al./em>. J Virol 2009; 83: 2216-2225.
HTLV-1 293T TransIT®-LT1 Lamb et al./em>. JBC 2009; 284: 6575-6584.
SIV/SK 293T/MDCK TransIT®-LT1 Masic et al./em>. J Gen Virol 2009; 90: 375-385.
MLV 293T TransIT®-293 Rulli et al./em>. J Virol 2008; 82: 6566-6575.
 

RNA Mediated Virus Production

TransIT-mRNA Transfection Kit Successfully Delivers Large Viral RNA

TransIT®-mRNA Transfection Kit Successfully Delivers Large Viral RNA. A 32 kb transcript of the murine coronavirus (MHV) was transfected into DBT cells using the TransIT®-mRNA Transfection Kit. Successful transfection was assessed 24-48 hours post-transfection by comparing a no RNA control (A) to the formation of syncytia (B) with MHV RNA. Syncytia were visualized by phase contrast microscopy.

Data courtesy of Mark Clementz, Loyola Univeristy of Chicago

TransIT-mRNA Transfection Kit Delivers RNA More Efficiently than Electroporation

TransIT®-mRNA Transfection Kit Delivers RNA More Efficiently than Electroporation. Huh7 cells were transfected with 2 µg of a yellow fever virus-luciferase expressing RNA replicon (YFV-luc) using the TransIT®-mRNA Transfection Kit or electroporation in PBS with 5 ug of the YFV-luc RNA replicon using an ECM630 Electroporator (BTX®) under optimal conditions. Twenty-four hours later, cells were harvested and assayed for luciferase activity and total protein content. Luciferase activity was normalized to total protein content to correct for differences in cell number between samples.

 

Citations for RNA mediated virus production using TransIT® Transfection Reagents

Virus Cell Type Product Citation
Picornavirus 293T/HeLa TransIT®-mRNA Greninger et al. J Virol 2012; 86: 3605-3616.
HCV HuH-7.5 TransIT®-mRNA Phan et al. J Virol; 2011; 85: 1193-1204.
Crimean-Congo Haemorrhagic Fever Virus BSR-T7/5 TransIT®-mRNA Bergeron et al. J Virol 2010; 84: 216-226.
HCV HuH-7 TransIT®-mRNA Jangra et al. J Virol 2010; 84: 6615-6625.
HEV PLC/PRF/5 TransIT®-mRNA Yamada et al. J Gen Virol 2009; 90: 457-462.

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Technical Resources

Mirus' Transfectopedia®: Why use recombinant viruses for nucleic acid delivery
Product Details: Find more information at the following product pages:

DNA Virus Production RNA Virus Production Baculovirus Production
TransIT®-VirusGEN™ Transfection Reagent
TransIT®-mRNA Transfection Kit TransIT®-Insect Transfection Reagent
TransIT®-Lenti Transfection Reagent
TransIT®-LT1 Transfection Reagent
TransIT®-293 Transfection Reagent
TransIT®-2020 Transfection Reagent

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Citation Database: Check if our reagents have been used by other researchers to transfect your cell type
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