CRISPR-Cas13 Inhibitors Block RNA Editing in Bacteria and Mammalian Cells.
Animals
Bacteria
/ genetics
Bacteriophages
/ genetics
CRISPR-Associated Proteins
/ antagonists & inhibitors
CRISPR-Cas Systems
/ genetics
Clustered Regularly Interspaced Short Palindromic Repeats
/ genetics
Escherichia coli
/ genetics
Gene Editing
HEK293 Cells
Humans
Leptotrichia
/ genetics
RNA
/ genetics
RNA Editing
/ genetics
AcrVIA
Cas13a
RNA editing
RNA targeting
anti-CRISPR
Journal
Molecular cell
ISSN: 1097-4164
Titre abrégé: Mol Cell
Pays: United States
ID NLM: 9802571
Informations de publication
Date de publication:
04 06 2020
04 06 2020
Historique:
received:
27
02
2020
revised:
10
03
2020
accepted:
25
03
2020
pubmed:
30
4
2020
medline:
20
9
2020
entrez:
30
4
2020
Statut:
ppublish
Résumé
Cas13 has demonstrated unique and broad utility in RNA editing, nucleic acid detection, and disease diagnosis; however, a constantly active Cas enzyme may induce unwanted effects. Bacteriophage- or prophage-region-encoded anti-CRISPR (acr) gene molecules provide the potential to control targeting specificity and potency to allow for optimal RNA editing and nucleic acid detection by spatiotemporally modulating endonuclease activities. Using integrated approaches to screen acrVI candidates and evaluate their effects on Cas13 function, we discovered a series of acrVIA1-7 genes that block the activities of Cas13a. These VI-A CRISPR inhibitors substantially attenuate RNA targeting and editing by Cas13a in human cells. Strikingly, type VI-A anti-CRISPRs (AcrVIAs) also significantly muffle the single-nucleic-acid editing ability of the dCas13a RNA-editing system. Mechanistically, AcrVIA1, -4, -5, and -6 bind LwaCas13a, while AcrVIA2 and -3 can only bind the LwaCas13-crRNA (CRISPR RNA) complex. These identified acr molecules may enable precise RNA editing in Cas13-based application and study of phage-bacterium interaction.
Identifiants
pubmed: 32348779
pii: S1097-2765(20)30225-2
doi: 10.1016/j.molcel.2020.03.033
pmc: PMC7299153
mid: NIHMS1587825
pii:
doi:
Substances chimiques
CRISPR-Associated Proteins
0
RNA
63231-63-0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
850-861.e5Subventions
Organisme : NIAID NIH HHS
ID : R03 AI097532
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI138203
Pays : United States
Organisme : NIGMS NIH HHS
ID : P20 GM113123
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI109317
Pays : United States
Organisme : NIGMS NIH HHS
ID : P20 GM103442
Pays : United States
Informations de copyright
Copyright © 2020 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests A provisional patent application pertaining to AcrVIA genes for CRISPR-related technologies and their applications has been filed by the University of North Dakota; Daping Hospital; and Army Medical University, Chongqing, China.
Références
Nucleic Acids Res. 2019 Jan 8;47(D1):D427-D432
pubmed: 30357350
Nat Commun. 2019 Aug 19;10(1):3728
pubmed: 31427601
Mol Cell. 2018 Apr 19;70(2):327-339.e5
pubmed: 29551514
Science. 2007 Mar 23;315(5819):1709-12
pubmed: 17379808
Mol Cell. 2019 Jan 17;73(2):264-277.e5
pubmed: 30503773
Mol Biol Evol. 2016 Jul;33(7):1870-4
pubmed: 27004904
Science. 2018 Oct 12;362(6411):156-157
pubmed: 30309933
Mol Cell. 2017 May 4;66(3):373-383.e3
pubmed: 28475872
Cell. 2018 Sep 20;175(1):212-223.e17
pubmed: 30241607
Cell. 2017 Jan 12;168(1-2):150-158.e10
pubmed: 28041849
Nature. 2014 Oct 30;514(7524):633-7
pubmed: 25174707
Nature. 2017 Jun 15;546(7658):436-439
pubmed: 28448066
Nat Microbiol. 2018 Apr;3(4):461-469
pubmed: 29507349
Science. 2016 Aug 5;353(6299):aaf5573
pubmed: 27256883
Nucleic Acids Res. 2011 Jul;39(Web Server issue):W347-52
pubmed: 21672955
Nature. 2015 Oct 1;526(7571):136-9
pubmed: 26416740
Nature. 2020 Jan;577(7791):572-575
pubmed: 31942067
Science. 2018 Apr 27;360(6387):439-444
pubmed: 29449508
Curr Opin Microbiol. 2017 Jun;37:67-78
pubmed: 28605718
Science. 2018 Aug 31;361(6405):866-869
pubmed: 30166482
Mol Cell. 2017 Feb 16;65(4):618-630.e7
pubmed: 28065598
Science. 2008 Dec 19;322(5909):1843-5
pubmed: 19095942
Nature. 2013 Jan 17;493(7432):429-32
pubmed: 23242138
iScience. 2019 Mar 29;13:55-68
pubmed: 30822746
Cell. 2017 Aug 10;170(4):714-726.e10
pubmed: 28757251
Methods. 2019 Mar 1;156:16-24
pubmed: 30502398
Cell. 2017 Sep 7;170(6):1224-1233.e15
pubmed: 28844692
Nature. 2016 Oct 13;538(7624):270-273
pubmed: 27669025
Nat Commun. 2018 Jul 25;9(1):2919
pubmed: 30046034
Nature. 2017 Oct 12;550(7675):280-284
pubmed: 28976959
J Vis Exp. 2013 Sep 16;(79):e50762
pubmed: 24084388
Cell. 2019 Oct 3;179(2):448-458.e11
pubmed: 31564454
Science. 2017 Nov 24;358(6366):1019-1027
pubmed: 29070703
Nat Struct Mol Biol. 2019 Apr;26(4):315-321
pubmed: 30936531
Mol Cell. 2018 Nov 1;72(3):404-412
pubmed: 30388409
Mol Cell. 2017 Jul 6;67(1):117-127.e5
pubmed: 28602637
Cell. 2016 Dec 15;167(7):1829-1838.e9
pubmed: 27984730
Nat Rev Microbiol. 2018 Jan;16(1):12-17
pubmed: 29062071
Nat Struct Mol Biol. 2016 Sep;23(9):868-70
pubmed: 27455460
Mol Biol Evol. 2004 Mar;21(3):587-98
pubmed: 14694080
Nat Microbiol. 2016 Jun 13;1(8):16085
pubmed: 27573108
Science. 2018 Oct 12;362(6411):240-242
pubmed: 30190308
mBio. 2018 Dec 4;9(6):
pubmed: 30514786
Cell. 2017 Jan 12;168(1-2):121-134.e12
pubmed: 28086085
Nat Commun. 2019 Mar 8;10(1):1136
pubmed: 30850590
Science. 2017 Apr 28;356(6336):438-442
pubmed: 28408723
Mol Cell. 2019 Jan 17;73(2):278-290.e4
pubmed: 30503774
Nat Biotechnol. 2013 Sep;31(9):822-6
pubmed: 23792628
Nature. 2019 Jun;570(7760):241-245
pubmed: 31142834
mBio. 2014 Apr 15;5(2):e00896
pubmed: 24736222
Mol Cell. 2015 Nov 5;60(3):385-97
pubmed: 26593719
Cell. 2018 Apr 19;173(3):665-676.e14
pubmed: 29551272
Science. 2018 Oct 12;362(6411):236-239
pubmed: 30190307
Nat Struct Mol Biol. 2019 Apr;26(4):308-314
pubmed: 30936526
Science. 2018 Apr 27;360(6387):444-448
pubmed: 29700266
Cell. 2015 May 21;161(5):1164-1174
pubmed: 25959775
Cell Host Microbe. 2019 Jun 12;25(6):815-826.e4
pubmed: 31155345
Science. 2019 Apr 19;364(6437):289-292
pubmed: 30819928