CRISPR-Cas9-based gene editing technology is a powerful new tool in biology. By simply altering the sequence of the single-guide RNA (sgRNA), one can reprogram Cas9 to target different sites in the genome with relative ease. Creative Biogene provides specialized and comprehensive sgRNA design and validation solutions for our clients' CRISPR/Cas9 experiments. Our scientists combine their extensive expertise with advanced industry technologies and are committed to creating optimized sgRNA sequences for our clients to meet their research demands.

Introduction of sgRNA Design

The performance of CRISPR/Cas relies on well-designed single-guide RNA (sgRNA). Since sgRNAs are responsible for recruiting Cas9 to specific genomic sites for targeted cleavage, the design of sgRNAs is critical to the success of gene editing experiments. The sgRNA is derived from the fusion of natural tracrRNA (transactivating crRNA) and crRNA (CRISPR RNA). Taking the widely used spCas9 system as an example, before designing sgRNAs, PAM sequences should be searched for near the target genome (PAM sequences are NGG, N=A/T/C/G), and gene-specific sgRNA template sequences are the neighboring motifs located in the sequence adjacent to the pre-interval region of the PAM sequence. The sgRNA, by directing the Cas 9 protein, can be sheared 3-4 bases upstream of the PAM sequence, inducing cells to undergo double-stranded breaks and form mutations in the target gene.