PasI (ER1861-200 units) restriction endonuclease was purchased from Thermo Fisher. dCTP, dCDP, dCMP, CTP, dUTP, dUMP, dTTP, dTMP, dGMP, GMP, dT, and dU were purchased from Beyotime (Shanghai, China), Aladdin (Shanghai, China), and Macklin (Shanghai, China). The sgRNA1 (5′-UAAUUUCUACUAAGUGUAGAUCCGCUUGCGGUGCCAUUAAU-3′, PAM: TTTG) and sgRNA2 (5′-UAAUUUCUACUAAGUGUAGAUUGGGCUUUGGCAAUUGCAUUCAU, PAM: TTTA-3′) for LbCas12a, and the sgRNA3 (5′-UGUGAUAAAUUCAUGAAGCGCGAGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUU-3′, PAM: TGG) and sgRNA4 (5′-AAAUAUCUUCGAUCCGUGGGGCUGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUU-3, PAM: GGG) for SpCas9 were synthesized by Tsingke Biotechnology. UV/UV–vis spectroscopic measurements were made using a NANODROP ONE (Thermo SCIENTIFIC), or using a Biotek Synergy 2 reader for 96-well plates.
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