Photoactivatable CRISPR/Cas9 System

A photoactivatable CRISPR/Cas9 system consisting of the Cas9 protein, synthetic 102-nt sgRNA or a pair of guide crRNA/tracrRNA, and blocking photocleavable oligodeoxyribonucleotides has been designed. Photocleavable oligodeoxyribonucleotides contain two or three photodegradable linkers in their structure. The thermal stability of complexes of these modified oligonucleotides with guide crRNA decreases with increasing the number of linkers. The kinetics of the photomodified oligodeoxyribonucleotides cleavage has been investigated. It has been shown that UV irradiation of the CRISPR/Cas9 system in the presence of photomodified blocking oligodeoxyribonucleotides can result in photoactivatable gene editing in vitro.