Cas-9 Dead mRNA-LNP

Cas9 (CRISPR-associated protein 9) is a dual RNA-guided DNA endonuclease associated with the clustered regularly interspaced short palindromic repeat (CRISPR) adaptive immune system originally discovered in Streptococcus pyogenes. The Cas9 protein is widely used as a genome engineering tool together with a small guide RNA (sgRNA) or gRNA to induce site-directed double-strand breaks or insertions in DNA, so-called CRISPR/Cas9 gene editing. CRISPR/Cas9 has been the basis for the development of numerous molecular systems used in research and, increasingly, in medical practice. Point mutations D10A and H840A alter 2 residues important for endonuclease activity of Cas9, resulting in Cas9 inactivation. The mutated Cas9 protein devoid of nucleolytic activity, known as Cas9 dead or dCas9, provide a technology for precise epigenetic DNA modification by delivering functional methylases and acetylases to genomic sites.
This product is designed as a tool for the delivery and expression of Cas9-dead mRNA for research. The product leverages the lipid nanoparticle (LNP) technology platform for simple and efficient delivery of Cas9-dead mRNA to a variety of mammalian cells in vitro and in vivo. The LNPs used are formulated with SM-102, DSPC, cholesterol and DMG-PEG2000 at an optimal molar concentration for a high rate of encapsulation and efficient mRNA delivery. Cas9-dead protein is 1368 amino acids, approximately 150 kD, and the GenPept accession number is QPF47903.