mRNA, viral RNA and CRISPR gRNA transfection kit: TransIT-mRNA

The TransIT®-mRNA Transfection Kit from Mirus Bio enables the highly efficient transfection of large RNA and CRISPR guide RNA into a broad range of cell types with minimal cellular toxicity. With no need for media changes with serum samples, each kit is supplied with the TransIT-mRNA Transfection Reagent and the mRNA Boost Reagent. 

Benefits of using the TransIT-mRNA Transfection Kit
• Low cellular toxicity
• High efficiency delivery
• Compatible with serum
• Deliver RNA of various sizes

Efficient genome editing with Cas9 mRNA and guide RNA pligonucleotides

HEK293T/17, U2OS and NHDF cells were co-transfected with 0.5 µg of Cas9 encoding mRNA, 5meC, ψ (Trilink Biotechnologies) and 25nM of PPIB targeting 2-part gRNA (Dharmacon) using TransIT-mRNA Transfection Kit (0.5 µl/well of 24-well plate of both mRNA Reagent and Boost, Mirus Bio). A T7E1 mismatch detection assay was used to measure cleavage efficiency at 48 hours post-transfection.

High efficiency and low toxicity transfection following 14 consecutive transfections with theTransIT-mRNA Transfection Kit

Repeated daily transfections were performed in the same population of BJ fibroblasts using TransIT-mRNA Transfection Kit, Lipofectamine® RNAiMAX (Life Technologies) & Stemfect™ RNA Transfection Kit (Stemgent) - with a capped & polyadenylated EGFP mRNA incorporating pseudouridine & 5mC modified bases (Trilink Biotechnologies, Inc.). Multiple reagent-to-RNA ratios were tested & the optimal ratio is represented. Transfections were performed in 12-well plates using the indicated reagent-to-RNA ratios to deliver 1 µg of RNA. GFP & phase contrast images were taken in the same field of view everyday after transfection. Transfection efficiency was measured by flow cytometry on a Guava® easyCyte™ 5HT following 14 consecutive daily transfections (blue bars). Cell viability was determined using cell counts measured during flow cytometry (black line grey bars). Error bars represent the standard deviation of triplicate wells. 

TransIT-mRNA Transfection Kit transfects GFP mRNA into DC 2.4 dendritic cells

Using TransIT-mRNA Transfection Kit, DC 2.4 cells were transfected with (A) 0.5 µg, (B) 1 µg and (C) 2.5 µg of capped and polyadenylated mRNA encoding GFP with 1 µl TransIT-mRNA Reagent and 1 µl Boost. Cells were seeded overnight at 100,000 cells/well in 24-well plates. Images were taken 10 hours post-transfection. Data courtesy of Kyle Phua (Principal Investigator: Kam W. Leong), Duke University. 

Applications
• CRISPR/CAS9 – genome editing 
• Virus production
• Protein expression

Testimonial
“Our lab recently used the TransIT-mRNA Transfection Kit to show that intracellular delivery of HPLC-purified and pseudouridine-containing mRNA can translate very efficiently without immune activation which is ideal for mRNA-based gene therapy applications. TransIT-mRNA further facilitated this work through low toxicity transfections of HEK 293T, human dendritic cells (DCs) and primary keratinocytes (Karikóe t al. Nucleic Acids Research, 39:e142, 2011).” 
Dr. Katalin Kariko, University of Pennsylvania, Department of Neurosurgery

Free samples
Request a free TransIT-mRNA Transfection Kit sample

Material available for download
TransIT-mRNA Transfection Kit protocol
TransIT-mRNA Transfection Kit protocol for CRISPR/Cas9 mRNA and gRNA delivery 
TransIT-mRNA Transfection Kit quick reference protocol

TransIT-mRNA citations

Lipofectamine is a registered trademark of Life Technologies , Inc.
Guava and easyCyte are registered trademarks of MERCK KGAA
Stemfect is a trademark of Stemgent, Inc.