Harnessing noncanonical crRNA for highly efficient genome editing

• DOI: 10.1038/s41467-024-48012-x
• Publication Date: 2024-05-07T16:01:52Z
• Authors: Guanhua Xun; Zhixin Zhu; Nilmani Singh; Jingxia Lu; Piyush K. Jain; Huimin Zhao
• Keywords: Trans-activating crRNA; Multiplex

The CRISPR-Cas12a system is more advantageous than the widely used CRISPR-Cas9 in terms of specificity and multiplexibility. However, its on-target editing efficiency typically much lower that system. Here we improved by simply incorporating 2-aminoadenine (base Z, which alters canonical Watson-Crick base pairing) into crRNA to increase binding affinity between complementary DNA target. resulting (named zCRISPR-Cas12a thereafter) shows an comparable but with off-target effects mammalian cells. In addition, can be for precise gene knock-in highly efficient multiplex genome editing. Overall, superior system, our simple engineering strategy may extended other CRISPR-Cas family members as well their derivatives.