TransIT®-mRNA Transfection Reagent for RNA Mediated Virus Production

Figure 1. TransIT-mRNA Outperforms Competitor RNA Transfection Reagents.
HuH7 cells were transfected with YF-R.Luc2A-RP RNA (1ug/well of 12 well plate) using the TransIT-mRNA Kit (Mirus Bio), DMRIE-C (Life Technologies) and TransMessenger™ (Qiagen) following the recommended protocols. Cells were harvested 24 hours later and assayed for luciferase activity according to manufacturer's protocol. DMRIE-C and TransMessenger reagents required transfections in the absence of serum with a media change 4-6 hours post-transfection.

Figure 2. 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

Figure 3. 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.

Figure 4. TransIT-mRNA Protocol Saves Time
The protocol for TransIT-mRNA is simple and saves time. RNA transfections with TransIT-mRNA are over 3 hours faster than TransMessenger^TM (Qiagen) and can save you over 2 hours compared to electroporation using standard buffers.