Difference between revisions of "Part:BBa K3982004"

Revision as of 11:20, 11 October 2021

CODE M sgRNA for targeting wildtype katG gene in Mycobacterium tuberculosis

This sequence encodes for the customised sgRNA and is used for targeting wildtype katG gene in Mycobacterium tuberculosis.

Usage and Biology

Cas14a1 is a miniature cas protein used in CRISPR/Cas technology. It is a RNA guided nuclease which cleaves ssDNA without any Protospacer Adjacent Motif (PAM) specificity i.e. PAM independent ssDNA cleavage. This enable high-fidelity Single-nucleotide polymorphism (SNP) genotyping. It is guided by a single guide RNA (sgRNA) which has two components - CRISPR RNA (crRNA) and Trans-activating crispr RNA (tracrRNA). crRNA is a 20bp nucleotide sequence complementary to the target DNA. tracrRNA is linked to the crRNA with a tetraloop that is responsible for cas14 activity. It plays a role in maturation of crRNA and has a stem-loop structure in itself that binds to the ribonuclease.

Sequence and Features