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. 2023 Apr 3;8(15):14113–14121. doi: 10.1021/acsomega.3c00790

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© 2023 The Authors. Published by American Chemical Society

Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).

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The CRISPR-Cas12a system is functional in Amycolatopsis sp. CCTCC NO: M2011265. (A) Structure of the pULcrRNAkm-Cas12a plasmid. Cas12a: Cas12a from Francisella novicida, driven by the constitutive promotor P_km; crRNA: the crRNA expression cassette driven by the constitutive promotor P_hsp60; pA-rep: the replicon used in CCTCC NO: M2011265; ColE1: the replicon used in E. coli; and Apr: apramycin resistance gene. (B) Effects of Cas gene expression on the growth of CCTCC NO: M2011265 determined by OD₆₀₀ measurement. Error bars (mean ± SD) were derived from triplicate samples. (C) Number of transformant colonies obtained with plasmid pULΔvdh (vdh guide sequence inserted in pULcrRNAkm-Cas12a) and control plasmid pULcrRNA-km-Cas12a (no vdh guide sequence). The data were obtained from three independent transformation experiments.