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. 2022 Feb 14;13:835040. doi: 10.3389/fmicb.2022.835040

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Copyright © 2022 Zhang, Sun, Lian, Wang, Jia, Leng, Chen, Bai, Meng, Cai and Tang.

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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Deletion of the nuclear localization signal (NLS) regulates Cas13d subcellular localization. (A) Summary of the constructs used in this study. Cas13d contains two NLSs that allow Cas13d to enter the nucleus. Removal of the NLS enables Cas13d to cleave the complementary target RNA within the cytoplasm but not the nucleus. (B) HEK293T cells were transfected with the CasRx expression vector or (C) CasRx-del-NLS, and then, IFA was performed at 24 hpi; the HA tag at the C-terminus of Cas13d was stained and labeled with Alexa Fluor 568-conjugated goat antibodies. Nuclei were labeled with DAPI. Scale bar = 2 μm.