Wild-type Cas9 mRNA is a stabilized non-immunogenic messenger RNA (mRNA) that has been designed to produce high expression level of wild-type Cas9 protein. It creates a double stranded break at a target site delineated by RNA guide sequences. This mRNA is produced by in vitro transcription, stabilized at the 5’ end by modified nucleotides capping and contain a poly(A) tail at the 3’ end. mRNACas9 is optimized to yield improved stability and performance and to lower the cellular innate immunity response.
Plasmids and viral vectors are traditionally used in CRISPR genome editing to express the required proteins. However, there is a risk to use DNA instead of mRNA: Double stranded DNA breaks catalyze insertion of DNA at the cut site. At some substantial frequency, the protein expression vectors can integrate, which can lead to continuous expression of the nuclease or a previously silent sequence.
NLS-Cre Recombinase mRNA is a capped and polyadenylated messenger RNA encoding Cre recombinase fused to a nuclear localization sequence (NLS). Cre recombinase is a tyrosine recombinase that catalyzes recombination between two loxP sites.
This mRNA is produced by in vitro transcription, stabilized at the 5’ end by modified nucleotides capping and contain a poly(A) tail at the 3’ end. CRE mRNA is optimized to yield improved stability and performance and to lower the cellular innate immunity response. Plasmids and viral vectors are traditionally used in CRISPR genome editing to express the required proteins. However, there is a risk to use DNA instead of mRNA: Double stranded DNA breaks catalyze insertion of DNA at the cut site. At some substantial frequency, the protein expression vectors can integrate, which can lead to continuous expression of the nuclease or a previously silent sequence.