Cas13d mRNA

Applications

The Cas13d MRNAs encode for the RNA-guided Cas13d endonuclease used to induce site-directed RNA degradation. Cas13d employs CRISPR-associated RNAs (crRNAs) that contain a customizable 22-nt spacer sequence that can direct the Cas13d protein to specific RNA molecules for targeted RNA degradation. The high catalytic activity of Cas13d in human cells provides a potential mechanism for targeting specific viral RNA genome degradation and viral gene expression inhibition (Aquino-Jarquin, 2018)(Nhan Huynh, 2020) (Timothy R. Abbott, 2020).

Cas13d mRNAs resemble fully matured mRNAs with 5’cap1 structure and 3’ polyA tail, therefore ready to be translated by the ribosome. mRNA transfection provides several advantages over plasmid DNA (pDNA) delivery. It does not require nuclear uptake for being expressed since translation of mRNA occurs directly into cytoplasm. Indeed, nuclear delivery (transport through nuclear membrane) is one the principal barriers for transfecting slow or non-dividing cells and consequently, mRNA transfection is particularly attractive for such purpose. This approach presents also the advantage of being non-integrative which is particularly appealing for stem cells, regenerative medicine or vaccine fields. Contrary to pDNA, mRNA cannot lead to genetic insertion causing mutations. Moreover, the protein expression from the mRNA is promoter-independent and faster than with DNA. For transfection we recommend RmesFect™ (#RM21000) and RmesFect™ Stem (#RS31000).