High Efficiency and Low Toxicity Transfection Following 14 Consecutive Transfections with TransIT®-mRNA Transfection Kit. Repeated daily transfections were performed in the same population of BJ fibroblasts using TransIT®-mRNA Transfection Kit, Lipofectamine® RNAiMAX (Life Technologies) and Stemfect™ RNA Transfection Kit (Stemgent) - with a capped and polyadenylated EGFP mRNA incorporating pseudouridine and 5mC modified bases (Trilink Biotechnologies, Inc.). Multiple reagent-to-RNA ratios were tested and 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 and phase contrast images were taken in the same field of view everyday after transfection. Representative images are shown at Days 1, 5, 9 and 14 post-transfection. Cells transfected with TransIT®-mRNA Transfection Kit exhibit the highest transfection efficiency with minimal cytotoxicity. Lipofectamine® RNAiMAX has low transfection efficiency, whereas Stemfect™ RNA Transfection kit displays significant cytotoxicity. Transfection efficiency and cell count measurements using flow cytometry are shown below.
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. (
Karikó et al. Nucleic Acids Research, 39:e142, 2011)
Dr. Katalin Kariko
University of Pennsylvania - Department of Neurosurgery
Don't See Your Cell Type? Consult Reagent Agent® Transfection Database
Citation Database: Check if our reagents have been used by other researchers to transfect your cell type
Technical Support: Speak directly with a transfection expert