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. 2022 Sep 2;12:978533. doi: 10.3389/fonc.2022.978533

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Copyright © 2022 Wu, Huo, Yang, Liu, Wu, Feng, Wang, Li, Jia, Zhang, Gao and Yu

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Preparation of CRISPR/Cas9-_3NLS/sgHMGA2 and verification of its cleavage activity (A) The Sanger sequencing result of CRISPR/Cas9-_3NLS; black lines indicate the bases encoding two inserted NLSs and one innate NLS, respectively; red line represents the stop code (TAA). (B) SDS-PAGE and Coomassie Brilliant blue staining for analyzing the induced expression and purification of CRISPR/Cas9-_3NLS. (C) The Sanger sequencing results of the sgHMGA2 expression plasmid; black lines indicate the bases encoding sgHMGA2-1, sgHMGA2-2, and sgHMGA2-3, respectively. (D) The nucleic acid electrophoresis results of the purified sgHMGA2-1, sgHMGA2-2, and sgHMGA2-3. (E, F) The nucleic acid electrophoresis results of the fragments obtained from cleaving artificially synthesized double-stranded DNA by CRISPR/Cas9-_3NLS/sgHMGA2; the length of DNA: 568 bp (E) and 418 bp (F).