Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex

Guo, Tai Wei; Bartesaghi, Alberto; Yang, Hui; Falconieri, Veronica; Rao, Prashant; Merk, Alan; Eng, Edward T.; Raczkowski, Ashleigh M.; Fox, Tara; Earl, Lesley A.; Patel, Dinshaw J.; Subramaniam, Sriram

doi:10.1016/j.cell.2017.09.006

Title: Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex

Abstract

Prokaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. Here, a central element of this immune system is an RNA-guided surveillance complex capable of targeting non-self DNA or RNA for degradation in a sequence- and site-specific manner analogous to RNA interference. Although the complexes display considerable diversity in their composition and architecture, many basic mechanisms underlying target recognition and cleavage are highly conserved. Using cryoelectron microscopy (cryo-EM), we show that the binding of target double-stranded DNA (dsDNA) to a type I-F CRISPR system yersinia (Csy) surveillance complex leads to large quaternary and tertiary structural changes in the complex that are likely necessary in the pathway leading to target dsDNA degradation by a trans-acting helicase-nuclease. Comparison of the structure of the surveillance complex before and after dsDNA binding, or in complex with three virally encoded anti-CRISPR suppressors that inhibit dsDNA binding, reveals mechanistic details underlying target recognition and inhibition.

Authors:

Guo, Tai Wei ^[1]; Bartesaghi, Alberto ^[1]; Yang, Hui ^[2]; Falconieri, Veronica ^[1]; Rao, Prashant ^[1]; Merk, Alan ^[1]; Eng, Edward T. ^[3]; Raczkowski, Ashleigh M. ^[3]; Fox, Tara ^[4]; Earl, Lesley A. ^[1]; Patel, Dinshaw J. ^[2]; Subramaniam, Sriram ^[1]

National Inst. of Health (NIH), Bethesda, MD (United States)
Memorial Sloan Kettering Cancer Center, New York, NY (United States)
New York Structural Biology Center, New York, NY (United States)
National Inst. of Health (NIH), Bethesda, MD (United States); Frederick National Lab. for Cancer Research, MD (United States)

Publication Date:: 2017-10-05

Research Org.:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Org.:: National Inst. of Health; Memorial Sloan Kettering Cancer Center; Center for Cancer Research, National Cancer Inst.; Simons Foundation; Agouron Inst.

OSTI Identifier:: 1430317

Grant/Contract Number:: HHSN261200800001E; P30CA008748; GM104962; 349247; GM103310; F00316; S10 Od019994-01

Resource Type:: Accepted Manuscript

Journal Name:: Cell

Additional Journal Information:: Journal Volume: 171; Journal Issue: 2; Journal ID: ISSN 0092-8674

Publisher:: Elsevier

Country of Publication:: United States

Language:: ENGLISH

Subject:: 59 BASIC BIOLOGICAL SCIENCES; CRISPR-Cas Csy complexes; CRISPR-Cas systems; adaptive immunity; PAM recognition; DNA-protein interactions; conformational transitions; gene editing; anti-CRISPR inhibitors; cryoelectron microscopy

Citation Formats


                    Guo, Tai Wei, Bartesaghi, Alberto, Yang, Hui, Falconieri, Veronica, Rao, Prashant, Merk, Alan, Eng, Edward T., Raczkowski, Ashleigh M., Fox, Tara, Earl, Lesley A., Patel, Dinshaw J., and Subramaniam, Sriram. Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex.  United States: N. p., 2017. 
Web.  doi:10.1016/j.cell.2017.09.006.

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                    Guo, Tai Wei, Bartesaghi, Alberto, Yang, Hui, Falconieri, Veronica, Rao, Prashant, Merk, Alan, Eng, Edward T., Raczkowski, Ashleigh M., Fox, Tara, Earl, Lesley A., Patel, Dinshaw J., & Subramaniam, Sriram. Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex.  United States.  https://doi.org/10.1016/j.cell.2017.09.006

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                    Guo, Tai Wei, Bartesaghi, Alberto, Yang, Hui, Falconieri, Veronica, Rao, Prashant, Merk, Alan, Eng, Edward T., Raczkowski, Ashleigh M., Fox, Tara, Earl, Lesley A., Patel, Dinshaw J., and Subramaniam, Sriram. Thu .  
"Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex".  United States.  https://doi.org/10.1016/j.cell.2017.09.006.  https://www.osti.gov/servlets/purl/1430317.

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@article{osti_1430317,

  title        = {Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex},

  author       = {Guo, Tai Wei and Bartesaghi, Alberto and Yang, Hui and Falconieri, Veronica and Rao, Prashant and Merk, Alan and Eng, Edward T. and Raczkowski, Ashleigh M. and Fox, Tara and Earl, Lesley A. and Patel, Dinshaw J. and Subramaniam, Sriram},

  abstractNote = {Prokaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. Here, a central element of this immune system is an RNA-guided surveillance complex capable of targeting non-self DNA or RNA for degradation in a sequence- and site-specific manner analogous to RNA interference. Although the complexes display considerable diversity in their composition and architecture, many basic mechanisms underlying target recognition and cleavage are highly conserved. Using cryoelectron microscopy (cryo-EM), we show that the binding of target double-stranded DNA (dsDNA) to a type I-F CRISPR system yersinia (Csy) surveillance complex leads to large quaternary and tertiary structural changes in the complex that are likely necessary in the pathway leading to target dsDNA degradation by a trans-acting helicase-nuclease. Comparison of the structure of the surveillance complex before and after dsDNA binding, or in complex with three virally encoded anti-CRISPR suppressors that inhibit dsDNA binding, reveals mechanistic details underlying target recognition and inhibition.},

  doi          = {10.1016/j.cell.2017.09.006},

  journal      = {Cell},

  number       = 2,

  volume       = 171,

  place        = {United States},

  year         = {2017},

  month        = {10}

}

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Inhibition mechanisms of AcrF9, AcrF8, and AcrF6 against type I-F CRISPR–Cas complex revealed by cryo-EM

Journal Article Zhang, Kaiming ; Wang, Shuo ; Li, Shanshan ; ... - Proceedings of the National Academy of Sciences of the United States of America

Prokaryotes and viruses have fought a long battle against each other. Prokaryotes use CRISPR–Cas-mediated adaptive immunity, while conversely, viruses evolve multiple anti-CRISPR (Acr) proteins to defeat these CRISPR–Cas systems. The type I-F CRISPR–Cas system in Pseudomonas aeruginosa requires the crRNA-guided surveillance complex (Csy complex) to recognize the invading DNA. Although some Acr proteins against the Csy complex have been reported, other relevant Acr proteins still need studies to understand their mechanisms. As such, here, we obtain three structures of previously unresolved Acr proteins (AcrF9, AcrF8, and AcrF6) bound to the Csy complex using electron cryo-microscopy (cryo-EM), with resolution at 2.57more »« less
Cited by 1
https://doi.org/10.1073/pnas.1922638117

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Broad-spectrum enzymatic inhibition of CRISPR-Cas12a

Journal Article Knott, Gavin J. ; Thornton, Brittney W. ; Lobba, Marco J. ; ... - Nature Structural & Molecular Biology

Cas12a is a bacterial RNA-guided nuclease used widely for genome editing and, more recently, as a molecular diagnostic. In bacteria, Cas12a enzymes can be inhibited by bacteriophage-derived proteins, anti-CRISPRs (Acrs), to thwart clustered regularly interspaced short palindromic repeat (CRISPR) adaptive immune systems. How these inhibitors disable Cas12a by preventing programmed DNA cleavage is unknown. We show that three such inhibitors (AcrVA1, AcrVA4 and AcrVA5) block Cas12a activity via functionally distinct mechanisms, including a previously unobserved enzymatic strategy. AcrVA4 and AcrVA5 inhibit recognition of double-stranded DNA (dsDNA), with AcrVA4 driving dimerization of Cas12a. In contrast, AcrVA1 is a multiple-turnover inhibitor thatmore »« less
Cited by 24
https://doi.org/10.1038/s41594-019-0208-z

Full Text Available
Mechanism of CRISPR-RNA guided recognition of DNA targets in Escherichia coli

Journal Article van Erp, Paul B. G. ; Jackson, Ryan N. ; Carter, Joshua ; ... - Nucleic Acids Research

In bacteria and archaea, short fragments of foreign DNA are integrated into Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) loci, providing a molecular memory of previous encounters with foreign genetic elements. In Escherichia coli, short CRISPR-derived RNAs are incorporated into a multi-subunit surveillance complex called Cascade (CRISPR-associated complex for antiviral defense). Recent structures of Cascade capture snapshots of this seahorse-shaped RNA-guided surveillance complex before and after binding to a DNA target. Here we determine a 3.2 Å x-ray crystal structure of Cascade in a new crystal form that provides insight into the mechanism of double-stranded DNA binding. Molecular dynamic simulationsmore »« less
Cited by 19
https://doi.org/10.1093/nar/gkv793

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Title: Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex

Abstract

Citation Formats

PHENIX: a comprehensive Python-based system for macromolecular structure solution journal, January 2010

Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease journal, July 2014

Structure of -galactosidase at 3.2-A resolution obtained by cryo-electron microscopy journal, July 2014

Bacteriophage genes that inactivate the CRISPR/Cas bacterial immune system journal, December 2012

Multiple mechanisms for CRISPR–Cas inhibition by anti-CRISPR proteins journal, September 2015

One number does not fit all: Mapping local variations in resolution in cryo-EM reconstructions journal, November 2013

The role of Cas8 in type I CRISPR interference journal, June 2015

Structure Reveals Mechanisms of Viral Suppressors that Intercept a CRISPR RNA-Guided Surveillance Complex journal, March 2017

Features and development of Coot journal, March 2010

Visualizing density maps with UCSF Chimera journal, January 2007

Csy4 relies on an unusual catalytic dyad to position and cleave CRISPR RNA: Mechanism of CRISPR RNA cleavage journal, April 2012

Structural basis for promiscuous PAM recognition in type I–E Cascade from E. coli journal, February 2016

CasA mediates Cas3-catalyzed target degradation during CRISPR RNA-guided interference journal, April 2014

DNA Targeting by a Minimal CRISPR RNA-Guided Cascade journal, September 2016

A Conserved Structural Chassis for Mounting Versatile CRISPR RNA-Guided Immune Responses journal, June 2015

Crystal structure of the CRISPR RNA-guided surveillance complex from Escherichia coli journal, August 2014

ePMV Embeds Molecular Modeling into Professional Animation Software Environments journal, March 2011

Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM journal, May 2013

EMAN: Semiautomated Software for High-Resolution Single-Particle Reconstructions journal, December 1999

Comparative genomics of defense systems in archaea and bacteria journal, March 2013

An updated evolutionary classification of CRISPR–Cas systems journal, September 2015

CRISPR-Cas immunity in prokaryotes journal, October 2015

The solution structure of an anti-CRISPR protein journal, October 2016

Diverse evolutionary roots and mechanistic variations of the CRISPR-Cas systems journal, August 2016

Crystal structure of a CRISPR RNA-guided surveillance complex bound to a ssDNA target journal, August 2014

Structures and mechanisms of CRISPR RNA-guided effector nucleases journal, April 2017

Crystal Structure of the CRISPR-Cas RNA Silencing Cmr Complex Bound to a Target Analog journal, May 2015

[20] Processing of X-ray diffraction data collected in oscillation mode book, January 1997

Structural Variation of Type I-F CRISPR RNA Guided DNA Surveillance journal, August 2017

Inactivation of CRISPR-Cas systems by anti-CRISPR proteins in diverse bacterial species journal, June 2016

Alternate binding modes of anti-CRISPR viral suppressors AcrF1/2 to Csy surveillance complex revealed by cryo-EM structures journal, June 2017

CTFFIND4: Fast and accurate defocus estimation from electron micrographs journal, November 2015

Mechanism of foreign DNA recognition by a CRISPR RNA-guided surveillance complex from Pseudomonas aeruginosa journal, February 2015

Directional R-Loop Formation by the CRISPR-Cas Surveillance Complex Cascade Provides Efficient Off-Target Site Rejection journal, March 2015

RELION: Implementation of a Bayesian approach to cryo-EM structure determination journal, December 2012

Protospacer recognition motifs: Mixed identities and functional diversity journal, February 2013

Automated molecular microscopy: The new Leginon system journal, July 2005

Direct observation of R-loop formation by single RNA-guided Cas9 and Cascade effector complexes journal, May 2014

EMAN2: An extensible image processing suite for electron microscopy journal, January 2007

Unravelling the structural and mechanistic basis of CRISPR–Cas systems journal, June 2014

Native Tandem and Ion Mobility Mass Spectrometry Highlight Structural and Modular Similarities in Clustered-Regularly-Interspaced Shot-Palindromic-Repeats (CRISPR)-associated Protein Complexes From Escherichia coli and Pseudomonas aeruginosa journal, November 2012

Mechanism of CRISPR-RNA guided recognition of DNA targets in Escherichia coli journal, August 2015

DoG Picker and TiltPicker: Software tools to facilitate particle selection in single particle electron microscopy journal, May 2009

Type I-E CRISPR-Cas Systems Discriminate Target from Non-Target DNA through Base Pairing-Independent PAM Recognition journal, September 2013

Structures of the RNA-guided surveillance complex from a bacterial immune system journal, September 2011

Biology and Applications of CRISPR Systems: Harnessing Nature’s Toolbox for Genome Engineering journal, January 2016

Structure Basis for Directional R-loop Formation and Substrate Handover Mechanisms in Type I CRISPR-Cas System journal, June 2017

Crystal Structure of Cpf1 in Complex with Guide RNA and Target DNA journal, May 2016

Crystal structure of the RNA-guided immune surveillance Cascade complex in Escherichia coli journal, August 2014

MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy journal, February 2017

New paradigm of functional regulation by DNA mimic proteins: Recent updates journal, December 2018

Cryo-EM structure of a type I-F CRISPR RNA guided surveillance complex bound to transposition protein TniQ journal, January 2020

Structural basis of DNA targeting by a transposon-encoded CRISPR–Cas system journal, December 2019

Expanding the mass range for UVPD-based native top-down mass spectrometry journal, January 2019

The interaction of phages and bacteria: the co-evolutionary arms race journal, December 2019

Keeping crispr in check: diverse mechanisms of phage-encoded anti-crisprs journal, May 2019

Anti‐CRISPR AcrIIC3 discriminates between Cas9 orthologs via targeting the variable surface of the HNH nuclease domain journal, August 2019

A CRISPR RNA Is Closely Related With the Size of the Cascade Nucleoprotein Complex journal, October 2019

Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble journal, July 2019

Anti‐CRISPRs: The natural inhibitors for CRISPR‐Cas systems journal, May 2019

Widespread anti-CRISPR proteins in virulent bacteriophages inhibit a range of Cas9 proteins journal, July 2018

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering journal, November 2019

Structure–function insights into the initial step of DNA integration by a CRISPR–Cas–Transposon complex journal, January 2020

Structural basis of a Tn7-like transposase recruitment and DNA loading to CRISPR-Cas surveillance complex journal, January 2020

DNA interference is controlled by R-loop length in a type I-F1 CRISPR-Cas system journal, June 2020

Type I-F CRISPR–Cas provides protection from DNA, but not RNA phages journal, October 2019

Structural organization of a Type III-A CRISPR effector subcomplex determined by X-ray crystallography and cryo-EM journal, February 2019

Structural insights into the inactivation of CRISPR-Cas systems by diverse anti-CRISPR proteins journal, March 2018

Structural insight into multistage inhibition of CRISPR-Cas12a by AcrVA4 journal, August 2019

Inhibition of CRISPR-Cas9 ribonucleoprotein complex assembly by anti-CRISPR AcrIIC2 journal, June 2019

Cryo-EM in drug discovery: achievements, limitations and prospects journal, June 2018

PAM identification by CRISPR-Cas effector complexes: diversified mechanisms and structures journal, September 2018

Csy4 relies on an unusual catalytic dyad to position and cleave CRISPR RNA: Mechanism of CRISPR RNA cleavage journal, April 2012

Expanding the mass range for UVPD-based native top-down mass spectrometry text, January 2019

Bacteriophage Cooperation Suppresses CRISPR-Cas3 and Cas9 Immunity journal, August 2018

Structure Reveals a Mechanism of CRISPR-RNA-Guided Nuclease Recruitment and Anti-CRISPR Viral Mimicry journal, April 2019

PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010

Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease
journal, July 2014

Structure of -galactosidase at 3.2-A resolution obtained by cryo-electron microscopy
journal, July 2014

Bacteriophage genes that inactivate the CRISPR/Cas bacterial immune system
journal, December 2012

Multiple mechanisms for CRISPR–Cas inhibition by anti-CRISPR proteins
journal, September 2015

One number does not fit all: Mapping local variations in resolution in cryo-EM reconstructions
journal, November 2013

The role of Cas8 in type I CRISPR interference
journal, June 2015

Structure Reveals Mechanisms of Viral Suppressors that Intercept a CRISPR RNA-Guided Surveillance Complex
journal, March 2017

Features and development of Coot
journal, March 2010

Visualizing density maps with UCSF Chimera
journal, January 2007

Csy4 relies on an unusual catalytic dyad to position and cleave CRISPR RNA: Mechanism of CRISPR RNA cleavage
journal, April 2012

Structural basis for promiscuous PAM recognition in type I–E Cascade from E. coli
journal, February 2016

CasA mediates Cas3-catalyzed target degradation during CRISPR RNA-guided interference
journal, April 2014

DNA Targeting by a Minimal CRISPR RNA-Guided Cascade
journal, September 2016

A Conserved Structural Chassis for Mounting Versatile CRISPR RNA-Guided Immune Responses
journal, June 2015

Crystal structure of the CRISPR RNA-guided surveillance complex from Escherichia coli
journal, August 2014

ePMV Embeds Molecular Modeling into Professional Animation Software Environments
journal, March 2011

Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM
journal, May 2013

EMAN: Semiautomated Software for High-Resolution Single-Particle Reconstructions
journal, December 1999

Comparative genomics of defense systems in archaea and bacteria
journal, March 2013

An updated evolutionary classification of CRISPR–Cas systems
journal, September 2015

CRISPR-Cas immunity in prokaryotes
journal, October 2015

The solution structure of an anti-CRISPR protein
journal, October 2016

Diverse evolutionary roots and mechanistic variations of the CRISPR-Cas systems
journal, August 2016

Crystal structure of a CRISPR RNA-guided surveillance complex bound to a ssDNA target
journal, August 2014

Structures and mechanisms of CRISPR RNA-guided effector nucleases
journal, April 2017

Crystal Structure of the CRISPR-Cas RNA Silencing Cmr Complex Bound to a Target Analog
journal, May 2015

[20] Processing of X-ray diffraction data collected in oscillation mode
book, January 1997

Structural Variation of Type I-F CRISPR RNA Guided DNA Surveillance
journal, August 2017

Inactivation of CRISPR-Cas systems by anti-CRISPR proteins in diverse bacterial species
journal, June 2016

Alternate binding modes of anti-CRISPR viral suppressors AcrF1/2 to Csy surveillance complex revealed by cryo-EM structures
journal, June 2017

CTFFIND4: Fast and accurate defocus estimation from electron micrographs
journal, November 2015

Mechanism of foreign DNA recognition by a CRISPR RNA-guided surveillance complex from Pseudomonas aeruginosa
journal, February 2015

Directional R-Loop Formation by the CRISPR-Cas Surveillance Complex Cascade Provides Efficient Off-Target Site Rejection
journal, March 2015

RELION: Implementation of a Bayesian approach to cryo-EM structure determination
journal, December 2012

Protospacer recognition motifs: Mixed identities and functional diversity
journal, February 2013

Automated molecular microscopy: The new Leginon system
journal, July 2005

Direct observation of R-loop formation by single RNA-guided Cas9 and Cascade effector complexes
journal, May 2014

EMAN2: An extensible image processing suite for electron microscopy
journal, January 2007

Unravelling the structural and mechanistic basis of CRISPR–Cas systems
journal, June 2014

Native Tandem and Ion Mobility Mass Spectrometry Highlight Structural and Modular Similarities in Clustered-Regularly-Interspaced Shot-Palindromic-Repeats (CRISPR)-associated Protein Complexes From Escherichia coli and Pseudomonas aeruginosa
journal, November 2012

Mechanism of CRISPR-RNA guided recognition of DNA targets in Escherichia coli
journal, August 2015

DoG Picker and TiltPicker: Software tools to facilitate particle selection in single particle electron microscopy
journal, May 2009

Type I-E CRISPR-Cas Systems Discriminate Target from Non-Target DNA through Base Pairing-Independent PAM Recognition
journal, September 2013

Structures of the RNA-guided surveillance complex from a bacterial immune system
journal, September 2011

Biology and Applications of CRISPR Systems: Harnessing Nature’s Toolbox for Genome Engineering
journal, January 2016

Structure Basis for Directional R-loop Formation and Substrate Handover Mechanisms in Type I CRISPR-Cas System
journal, June 2017

Crystal Structure of Cpf1 in Complex with Guide RNA and Target DNA
journal, May 2016

Crystal structure of the RNA-guided immune surveillance Cascade complex in Escherichia coli
journal, August 2014

MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy
journal, February 2017

New paradigm of functional regulation by DNA mimic proteins: Recent updates
journal, December 2018

Cryo-EM structure of a type I-F CRISPR RNA guided surveillance complex bound to transposition protein TniQ
journal, January 2020

Structural basis of DNA targeting by a transposon-encoded CRISPR–Cas system
journal, December 2019

Expanding the mass range for UVPD-based native top-down mass spectrometry
journal, January 2019

The interaction of phages and bacteria: the co-evolutionary arms race
journal, December 2019

Keeping crispr in check: diverse mechanisms of phage-encoded anti-crisprs
journal, May 2019

Anti‐CRISPR AcrIIC3 discriminates between Cas9 orthologs via targeting the variable surface of the HNH nuclease domain
journal, August 2019

A CRISPR RNA Is Closely Related With the Size of the Cascade Nucleoprotein Complex
journal, October 2019

Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble
journal, July 2019

Anti‐CRISPRs: The natural inhibitors for CRISPR‐Cas systems
journal, May 2019

Widespread anti-CRISPR proteins in virulent bacteriophages inhibit a range of Cas9 proteins
journal, July 2018

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering
journal, November 2019

Structure–function insights into the initial step of DNA integration by a CRISPR–Cas–Transposon complex
journal, January 2020

Structural basis of a Tn7-like transposase recruitment and DNA loading to CRISPR-Cas surveillance complex
journal, January 2020

DNA interference is controlled by R-loop length in a type I-F1 CRISPR-Cas system
journal, June 2020

Type I-F CRISPR–Cas provides protection from DNA, but not RNA phages
journal, October 2019

Structural organization of a Type III-A CRISPR effector subcomplex determined by X-ray crystallography and cryo-EM
journal, February 2019

Structural insights into the inactivation of CRISPR-Cas systems by diverse anti-CRISPR proteins
journal, March 2018

Structural insight into multistage inhibition of CRISPR-Cas12a by AcrVA4
journal, August 2019

Inhibition of CRISPR-Cas9 ribonucleoprotein complex assembly by anti-CRISPR AcrIIC2
journal, June 2019

Cryo-EM in drug discovery: achievements, limitations and prospects
journal, June 2018

PAM identification by CRISPR-Cas effector complexes: diversified mechanisms and structures
journal, September 2018

Csy4 relies on an unusual catalytic dyad to position and cleave CRISPR RNA: Mechanism of CRISPR RNA cleavage
journal, April 2012

Expanding the mass range for UVPD-based native top-down mass spectrometry
text, January 2019

Bacteriophage Cooperation Suppresses CRISPR-Cas3 and Cas9 Immunity
journal, August 2018

Structure Reveals a Mechanism of CRISPR-RNA-Guided Nuclease Recruitment and Anti-CRISPR Viral Mimicry
journal, April 2019