The GenCrispr sgRNA synthesis kit is designed to synthesize gRNAs in vitro.
In the bacterial CRISPR/Cas9 system, the Cas9 nuclease associates with two RNAs, the CRISPR RNA (crRNA) and the trans-activating crRNA (tracrRNA), to direct sequence-specific DNA cleavage. The gRNA (guide RNA) is a fusion of the natural crRNA and tracrRNA components. It contains an 18-20 base variable sequence that can be changed to target any DNA sequence that is adjacent to an NGG proto-spacer adjacent motif (PAM) on the 3’ end of the target sequence.
The GenCrispr sgRNA synthesis kit allows user to fuse their choice of 18-20 base target sequence within the gRNA DNA sequence provided, by PCR fusion. The kit also provides materials for in-vitro transcription of the generated gRNA DNA in order to generate gRNAs which can be directly used for in-vivo genome editing.