Mixed Messages: a SNiP Double Feature

In each SNiP or “Small News in Pieces” we highlight standout research articles we’ve come across that feature Mirus Bio products… in bite-size pieces. Our eyes are always on the lookout for Mirus products in novel applications, and when choosing the topic for today’s post, we stumbled across not one, but two unique studies to share! In both studies discussed today, Mirus’ TransIT®-mRNA Transfection Kit was pivotal in establishing an mRNA delivery workflow for some unique recipients: whole live organisms!

The TransIT®-mRNA Transfection Kit was originally developed for transfecting cells in culture. It is highly efficient, serum compatible and suitable for delivering diverse species of RNA. For decades, TransIT®-mRNA has been used for a variety of applications such as virus production, protein expression and CRISPR genome-editing. But, usually in vitro. We were thus delighted to learn about two new and innovative applications of TransIT®-mRNA in vivo.

Our first highlight today comes from Moskowitzova et al., who used the TransIT®-mRNA Transfection Kit to condense and deliver luciferase mRNA to rat fetuses via intra-amniotic injections. The amniotic cavity surrounds fetuses growing in utero. Goals of this study included affirming that intra-amniotic delivery of lipopolyplexed mRNA is possible, in addition to determining the route of fetal uptake. In addition to swallowing and aspiration, the study’s results suggest that the transfection complexes spread via the bloodstream through gestational membranes, the umbilical cord and the placenta. By use of the TransIT®-mRNA Transfection Kit, the authors were able to contribute to our collective understanding of transamniotic nucleic acid therapy, a blossoming field of study.

Our second highlight comes from Weill et al., who developed a method for transfecting mRNA in planarians. Schmidtea mediterranea are a kind of planarian flatworm used as a model organism in regenerative medicine research owing to its remarkable ability to regenerate almost all lost body parts! Despite the significance of planarians to biopharmaceutical research, methods to transfect and study their genes are not plentiful. The authors outline a readily accessible method for transfecting planarians and hope that their work will contribute to further development of additional transgenic techniques for this model organism.

Read the full articles to learn more!

Citations

Title: Hematogenous Routing of Exogenous Messenger RNA Delivered Into the Amniotic Fluid
Authors: Kamila Moskowitzova, Ashlyn Whitlock et al.
Journal: JSR, Volume 289, Apr 2023.
DOI: 10.1016/j.jss.2023.03.037
Product Usage: The TransIT®-mRNA Transfection Kit was used to form firefly luciferase mRNA complexes in a 50 µl volume of PBS. The amniotic cavities of Sprague-Dawley dams were injected with the full volume of transfection complexes on gestational day 17. The presence of luciferase was assayed from fetuses harvested from daily timepoints from gestational day 18 to day 21.

Title: mRNA Transfection of S. mediterranea for Luminescence Analysis
Authors: Uri Weill, Richard Nelson Hall et al.
Journal: MIMB, Volume 2680, Jul 2023.
DOI: 10.1007/978-1-0716-3275-8_17
Product Usage: The TransIT®-mRNA Transfection Kit was used to form nanoluciferase mRNA complexes in a 25 µL volume. The entire volume was added to ~200,000 dissociated planarian cells in a 96-well plate well. For live worm transfection, 8 µL of the complexes was microinjected per worm. Luminescence was assessed by plate reader or microscopy and detected as soon as 4 hours post-transfection.

Read more articles featuring Mirus reagents in our Citations Database!