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. 2021 Dec 16;9:786354. doi: 10.3389/fchem.2021.786354

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Copyright © 2021 Qiao, Wu, Zhang, Luo, Wang and Ming.

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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NIR-I/CRISPR-Cas9-based imaging systems. (A) CRISPR-mediated endogenous labeling of proteins could combine with NIR dyes, and show NIR imaging following by excitation. CRISPR-Cas9 is used to knockin SNAP-tagged or Halo-tagged protein from its genomic locus. NIR dye bind with SNAP-tagged or Halo-tagged protein to give image. (B) Structures of the probes used for two-color imaging. (C) Western blots of CRISPR-Cas9 mediated tagging of vimentin with SNAP Tag or Halo Tag show a higher expression. (D) Confocal images of vimentin-SNAP knock-in cells labeled with 610CP-tubulin2 and 680SiR-SNAP probes. (E) Upper row, confocal images of vimentin-SNAP stained with 680SiR-SNAP, microtubule cytoskeleton stained with 610CP-tubulin2, and merge. Lower row, the corresponding STED images (reproduced from (Butkevich et al., 2018) with permission from ACS chemical biology).