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Drug Inducible CRISPR/Cas Systems

Zhang, Jingfang, Chen, Li, Zhang, Ju, Wang, Yu·2019·Computational and Structural Biotechnology Journal
DOI 10.1016/j.csbj.2019.07.015PMC PMC6709367
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Graphical abstract CC BY-NC-ND · PubMed Central
Fig. 1 CRISPR/Cas9 systems of drug induction at the transcriptional level. A. Tet based CRISPR/Cas9 systems for doxycycline regulation of Cas9 (left) or sgRNA (right) expression. AD, activation domain. B. Cre-ERT2-dependent CRISPR/Cas9 system. Cre-ERT2 removes the loxp-stop-loxp cassette and drives Cas9 expression dependent on 4-OHT regulated nuclear translocation.
Fig. 1 CRISPR/Cas9 systems of drug induction at the transcriptional level. A. Tet based CRISPR/Cas9 systems for doxycycline regulation of Cas9 (left) or sgRNA (right) expression. AD, activation domain. B. Cre-ERT2-dependent CRISPR/Cas9 system. Cre-ERT2 removes the loxp-stop-loxp cassette and drives Cas9 expression dependent on 4-OHT regulated nuclear translocation.CC BY-NC-ND · PubMed Central
Fig. 2 CRISPR/Cas9 systems of drug induction at the posttranslational level. A. Chemically induced proximity (CIP) systems. Drug induces split Cas9 dimerization and forming complete and functional Cas9 (upper panel); similarly, drug induces dimerization of dCas9 and transcriptional activation domains (ADs) for gene activation (lower panel). NES, nuclear export sequence; NLS, nuclear localization sequence. B. Intein splicing CRISPR/Cas9 system. C. Systems based on 4-OHT driven ER nuclear translocation. D. Allosterically regulated Cas9. E. Destabilized domains (DDs) controlled CRISPR/Cas9 systems. Small molecule stabilizes DD-Cas9 (left) or DD-PP7-ADs (right) and renders them drug inducible. DSB, double strand break.
Fig. 2 CRISPR/Cas9 systems of drug induction at the posttranslational level. A. Chemically induced proximity (CIP) systems. Drug induces split Cas9 dimerization and forming complete and functional Cas9 (upper panel); similarly, drug induces dimerization of dCas9 and transcriptional activation domains (ADs) for gene activation (lower panel). NES, nuclear export sequence; NLS, nuclear localization sequence. B. Intein splicing CRISPR/Cas9 system. C. Systems based on 4-OHT driven ER nuclear translocation. D. Allosterically regulated Cas9. E. Destabilized domains (DDs) controlled CRISPR/Cas9 systems. Small molecule stabilizes DD-Cas9 (left) or DD-PP7-ADs (right) and renders them drug inducible. DSB, double strand break.CC BY-NC-ND · PubMed Central
Fig. 3 Schematic presentation of two orthogonal CRISPR/Cas9 systems. A. Two independent chemically induced proximity (CIP) systems in combination with spCas9 and saCas9 respectively form an orthogonal CRISPR/Cas9 system for independent regulation of two genes. KRAB, Krüppel-associated box; VPR, VP64-P65-Rta. B. Two independent destabilized domain (DD) systems in combination with distinct aptamers constitute another orthogonal system. PH, P65-HSF1.
Fig. 3 Schematic presentation of two orthogonal CRISPR/Cas9 systems. A. Two independent chemically induced proximity (CIP) systems in combination with spCas9 and saCas9 respectively form an orthogonal CRISPR/Cas9 system for independent regulation of two genes. KRAB, Krüppel-associated box; VPR, VP64-P65-Rta. B. Two independent destabilized domain (DD) systems in combination with distinct aptamers constitute another orthogonal system. PH, P65-HSF1.CC BY-NC-ND · PubMed Central

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Cited by

Metadata

Authors
Zhang, J., Chen, L., Zhang, J., & Wang, Y.
Affiliations
  • School of Life Sciences, Beijing University of Chinese Medicine, Beijing 100029, China
  • State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
Venue
Computational and Structural Biotechnology Journal
Publisher
American Association for the Advancement of Science (AAAS)
Published
2019-01
Volume
17
Pages
1171-1177
ISSN
2001-0370
DOI
10.1016/j.csbj.2019.07.015
PMC
PMC6709367
Cited by
69
References
11
Language
en
Licence
Funders
  • National Basic Research Program of China (2015CB964800)
  • National Natural Science Foundation of China (31701280, 31571514)
  • Chinese Academy of Sciences (XDA16010504)

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