According to a genetic editing study published online by the British journal Nature on the 12th, a team of European and American scientists reported that the high-efficiency detection system for the genome-wide off-target effect of the CRISPR-Cas9 genome was tested in mice. The results of this research will facilitate the transformation of genome editing from research to clinical.

The CRISPR-Cas9 genome editing technology is known as the “genetic devil” and is considered to be an innovative technology with great potential in the history of human medicine, but only when it fully recognizes and eliminates additional non-target (off-target) mutations. Achieve clinical application.

The so-called off-target mutation refers to the attachment of the CRISPR-Cas9 complex to multiple sites in the genome and the erroneous DNA fragments are cleaved. Systems that detect this mutation must be tested in vitro as well as throughout the organism.

This time, the Massachusetts General Hospital and researchers at AstraZeneca, Sweden, described a highly sensitive "verification of in vivo off-targets" (VIVO) system that can detect CRISPR-Cas9. Off-target mutations in organisms. The VIVO system identifies potential off-target sites in advance and confirms that any of these sites have changed after genome editing. The team tested the accuracy of the system in mouse liver by designing a variety of guide RNAs (gRNAs) that target the Pcsk9 gene in mice, including promiscuous (targeting multiple sites) gRNAs and specificities. Higher gRNA.

The results confirmed that the VIVO system not only detected dozens of off-target mutations induced by promiscuous gRNA (including mutations with a rate of only 0.13%), but the correctly designed gRNA did not bring any detectable off-target mutations.

The researchers believe that the VIVO system sets an important standard for defining the off-target effects of genome editing in practical applications and confirms the importance of designing highly specific gRNAs. (Reporter Zhang Mengran)

Source: Technology Daily

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