On December 15th, 2016, Molecular Cell magazine published the latest research results of the CRISPR-Cas system by Wang Yanli from the Institute of Biophysics of the Chinese Academy of Sciences. The article is titled “C2c1-sgRNA Complex Structure Reveals RNA-Guided DNA Cleavage Mechanismâ€.
The CRISPR-Cas system consists of clustered and regularly spaced short palindromic repeats and its associated protein (Cas), an RNA-mediated adaptive immune system in prokaryotes whose primary function is to resist foreign invasion. Genetic material. In addition, the CRISPR-Cas system is widely used for genome editing, and CRISPR-Cas9 is the most widely used. Another recently discovered Cas protein, C2c1, has a similar function as Cas9. This can not help but make people think: Can C2c1 be developed into a new gene editing tool? The work of Wang Yanli's research group may provide some new theoretical foundations for the realization of this idea.
In this study, the panelists analyzed the crystal structure of C2c1 bound to sgRNA. This structure shows that C2c1 is composed of two regions, an REC region having a function of recognizing sgRNA and a NUC region having nuclease function. The sgRNA is artificially chimeric by crRNA and tracrRNA, wherein the crRNA binds to the central channel of C2c1 and the tracrRNA is placed in the groove of the outer surface of C2c1. Interestingly, by further observation, the researchers found that the sgRNA corresponding to C2c1 exhibited a structure that was significantly different from the sgRNA corresponding to Cas9 or the crRNA corresponding to the recently reported Cpf1. Upon the suggestion of this new structure, the researchers analyzed the sgRNA structure corresponding to C2c1 of other species on the one hand, and shortened the length of the sgRNA on the other hand, and carried out activity experiments. Both of these results preliminarily verified this uniqueness. The authenticity and effectiveness of the structure. In addition, the study also found that a single base mutation in the sequence of interest can significantly reduce C2c1 splicing activity, indicating that C2c1 has extremely stringent requirements for the targeted sequence of interest, and the results of this study will help develop new genome editing tools. Reduce the off-target phenomenon during gene editing.
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