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The CRISPR-Cas system consists of clusters of regularly spaced short echo repetition sequences and its associated proteins (Cas), which are RNA-mediated accessary immune systems in primary nuclear organisms and whose primary function is to protect against foreign invasions of genetic material.
addition, CRISPR-Cas systems are widely used in genome editing, with CRISPR-Cas9 being used most widely.
C2c1, another recently discovered Cas protein, has a function similar to Cas9.
this makes one wonder: Can C2c1 also be developed as a new gene editing tool? This work by Wang Yanli's team may provide some new theoretical basis for the realization of this idea.
study, team members analyzed the structure of C2c1 crystals combined with sgRNA.
the structure shows that C2c1 consists of two regions, the REC region with sgRNA recognition function and the NUC region with nuclease function.
sgRNA is artificially embedded with crRNA and tracerRNA, where crRNA binds to the central hole of C2c1 and tracrRNA is placed in a groove on the outer surface of C2c1.
interestingly, on closer inspection, the team found that the sgRNA corresponding to C2c1 showed a significantly different structure than the sgRNA corresponding to Cas9 or the crRNA corresponding to the recently reported CF1.
The new structure was prompted by researchers analyzing the sgRNA structure corresponding to C2c1 in other species, shortening the length of the sgRNA and conducting active experiments, both of which initially verified the authenticity and effectiveness of this unique structure.
In addition, the study also found that a single base mutation in the target sequence significantly reduced C2c1 shear activity, indicating that C2c1 has extremely strict requirements for targeted target sequences, and this finding could help develop new genome editing tools to reduce off-targeting in gene editing.
Liu Liang (postdoctoral) and Chen Peng (graduate student) of Wang Yanli's research group are the first and second authors of this paper, respectively, and the research has been funded by the Ministry of Science and Technology, the National Natural Science Foundation of China and the Strategic Pilot Science and Technology Special (Class B) of the Chinese Academy of Sciences.
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