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Lb2Cas12a and its engineered variants mediate genome editing in human cells

Cas12a‐mediated targeted genome engineering strategies have enabled a broad range of research and clinical applications. However, the limited target‐selection spectrum and low activity/fidelity remain a bottleneck for its widespread application in precision site‐specific human genome editing. Therefore, there exists an acute need to identify novel Cas12a nucleases with improved features for genome editing. By screening a range of candidate Cas12a nucleases, here we demonstrate that Lb2Cas12a possesses genome editing activity in human cells and it has greater flexibility in PAM (5ʹ‐BYYV‐3ʹ) selection. Furthermore, we engineered Lb2Cas12a to generate variants (Lb2Cas12a‐RVR and Lb2Cas12a‐RR), which greatly expands the target‐selection spectrum. Our study illustrated that Lb2Cas12a could be harnessed as additional genome editing tool for the manipulation of human genome.