HyperScribe™ One-Step sgRNA Synthesis Kit

The CRISPR/Cas9 system is a widely used genome editing technology, which uses Cas9 nuclease to cut specific sites of targeted sequences of genomic DNA under the guidance of sgRNA, also known as gRNA (guide RNA). gRNA is a fusion product consisting of 18-20 bp CRISPR RNA (crRNA) sequences that complement the target gene sequences, and Trans-activating crRNA (tracrRNA) sequences that bind to Cas9 nucleases.In the cell, gRNA bound to Cas9 nuclease and guided Cas9 nuclease to cut the target gene at the upstream position of the proto-spacer adjacent motif (PAM) sequence, causing frameshift mutations in the target gene locus and eventually resulting in deletion mutations of the target gene.

This kit is a One-step method for the synthesis of sgRNA based on PCR amplification and in vitro transcription dependent on T7 RNA polymerase. The kit provides a sgRNA reaction mix, including a Scaffold Template sequence and a downstream primer for amplification of the sequence. Users only need to design and synthesize the Target-specific DNA oligo (i.e. the upstream primer) to obtain the corresponding sgRNA through this kit. After purification, the synthesized sgRNA can be co-transfected with Cas9 mRNA (No.R1015), or gene edited in cells expressing Cas9 protein, or mixed with Cas9 nuclease for enzyme digestion identification experiment. The one-step method for synthesizing sgRNA is simpler to operate compared to the Two-Step method, However, the yield is lower than that of the two-step method. If the two-step method for synthesizing sgRNA is required, it is recommended to use the HyperScribe™ two-step sgRNA Synthesis Kit (No. K1421).