CRISPR-Cas Enzymes

CRISPR-Cas Enzymes

1st Edition - January 25, 2019

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  • Editor: Scott Bailey
  • eBook ISBN: 9780128167618
  • Hardcover ISBN: 9780128167601

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CRISPR-Cas Enzymes, Volume 616, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Topics covered in this release include CRISPR bioinformatics, A method for one-step assembly of Class 2 CRISPR arrays, Biochemical reconstitution and structural analysis of ribonucleoprotein complexes in Type I-E CRISPR-Cas systems, Mechanistic dissection of the CRISPR interference pathway in Type I-E CRISPR-Cas system, Site-specific fluorescent labeling of individual proteins within CRISPR complexes, Fluorescence-based methods for measuring target interference by CRISPR-Cas systems, Native State Structural Characterization of CRISRP Associated Complexes using Mass Spectrometry, and more.

Key Features

  • Provides the authority and expertise of leading contributors from an international board of authors
  • Presents the latest release in the Methods in Enzymology series
  • Updated release includes the latest information on the CRISPR-Cas Enzymes


Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

Table of Contents

  • 1. Predicting and visualizing features of CRISPR–Cas systems
    Matthew A. Nethery and Rodolphe Barrangou
    2. Reconstitution and biochemical characterization of ribonucleoprotein complexes in Type I-E CRISPR–Cas systems
    Yibei Xiao and Ailong Ke
    3. Sortase-mediated fluorescent labeling of CRISPR complexes
    Kaylee E. Dillard, Jeffrey M. Schaub, Maxwell W. Brown, Fatema A. Saifuddin, Yibei Xiao, Erik Hernandez, Samuel D. Dahlhauser, Eric V. Anslyn, Ailong Ke and Ilya J. Finkelstein
    4. Fluorescence-based methods for measuring target interference by CRISPR–Cas systems
    Phong T. Phan, Michael Schelling, Chaoyou Xue and Dipali G. Sashital
    5. Probing Cascade complex composition and stability using native mass spectrometry techniques
    Angela Patterson, Monika Tokmina-Lukaszewska and Brian Bothner
    6. High-throughput determination of in vivo DNA sequence preferences for Cas protein binding using Library-ChIP
    Joseph T. Wade
    7. Live-cell single-particle tracking photoactivated localization microscopy of Cascade-mediated DNA surveillance
    Bartosz Turkowyd, Hanna Müller-Esparza, Vanessa Climenti, Niklas Steube, Ulrike Endesfelder and Lennart Randau
    8. In vitro assembly of thermostable Csm complex in CRISPR–Cas type III/A system
    Kwang-Hyun Park, Yan An and Eui-Jeon Woo
    9. Investigation of the cyclic oligoadenylate signaling pathway of type III CRISPR systems
    Christophe Rouillon, Januka S. Athukoralage, Shirley Graham, Sabine Grüschow and Malcolm F. White
    10. A pipeline for characterization of novel Cas9 orthologs
    Tautvydas Karvelis, Joshua K. Young and Virginijus Siksnys
    11. Preparation and electroporation of Cas12a/Cpf1-guide RNA complexes for introducing large gene deletions in mouse embryonic stem cells
    Lucas Kissling, Asun Monfort, Daan C. Swarts, Anton Wutz and Martin Jinek
    12. Directed evolution studies of a thermophilic Type II-C Cas9
    Travis H. Hand, Anuska Das and Hong Li
    13. Kinetic characterization of Cas9 enzymes
    Mu-Sen Liu, Shanzhong Gong, Helen-Hong Yu, David W. Taylor and Kenneth A. Johnson
    14. Single-molecule FRET studies of Cas9 endonuclease
    Viktorija Globyte and Chirlmin Joo
    15. CRISPR–Cas molecular beacons as tool for studies of assembly of CRISPR–Cas effector complexes and their interactions with DNA
    Vladimir Mekler, Konstantin Kuznedelov, Leonid Minakhin, Karthik Murugan, Dipali G. Sashital and Konstantin Severinov
    16. Adapting dCas9-APEX2 for subnuclear proteomic profiling
    Xin D. Gao, Tomás C. Rodríguez and Erik J. Sontheimer
    17. CRISPR–Cas10 assisted editing of virulent staphylococcal phages
    S.M. Nayeemul Bari and Asma Hatoum-Aslan
    18. Reconstitution of CRISPR adaptation in vitro and its detection by PCR
    Robert D. Fagerlund, Timothy J. Ferguson, Howard W.R. Maxwell, Helen K. Opel-Reading, Kurt L. Krause and Peter C. Fineran

Product details

  • No. of pages: 451
  • Language: English
  • Copyright: © Academic Press 2019
  • Published: January 25, 2019
  • Imprint: Academic Press
  • eBook ISBN: 9780128167618
  • Hardcover ISBN: 9780128167601

About the Serial Volume Editor

Scott Bailey

Scott Bailey is at Johns Hopkins Bloomberg School of Public Health, USA

Affiliations and Expertise

Johns Hopkins Bloomberg School of Public Health, USA

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