Coverart for item
The Resource CRISPR-Cas Enzymes, edited by Scoll Bailey

CRISPR-Cas Enzymes, edited by Scoll Bailey

Label
CRISPR-Cas Enzymes
Title
CRISPR-Cas Enzymes
Statement of responsibility
edited by Scoll Bailey
Contributor
Editor
Subject
Language
eng
Summary
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
Member of
Cataloging source
N$T
Dewey number
572.86
Index
index present
LC call number
QH442
Literary form
non fiction
Nature of contents
  • dictionaries
  • bibliography
http://library.link/vocab/relatedWorkOrContributorName
Bailey, Scott
Series statement
Methods in enzymology,
Series volume
volume 616
http://library.link/vocab/subjectName
  • Genetic engineering
  • CRISPR (Genetics)
  • Genomics
  • Endonucleases
  • Restriction enzymes, DNA
  • CRISPR-Associated Protein 9
  • CRISPR-Cas Systems
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Gene Editing
  • SCIENCE
  • TECHNOLOGY & ENGINEERING
  • CRISPR (Genetics)
  • Endonucleases
  • Genetic engineering
  • Genomics
  • Restriction enzymes, DNA
Label
CRISPR-Cas Enzymes, edited by Scoll Bailey
Instantiates
Publication
Antecedent source
unknown
Bibliography note
Includes bibliographical references at the end of each chapter and indexes
Carrier category
online resource
Carrier category code
  • cr
Carrier MARC source
rdacarrier
Color
multicolored
Content category
text
Content type code
  • txt
Content type MARC source
rdacontent
Contents
  • Front Cover; CRISPR-Cas Enzymes; Copyright; Contents; Contributors; Preface; Chapter One: Predicting and visualizing features of CRISPR-Cas systems; 1. Introduction; 2. Identify and characterize CRISPR-Cas system enzymes; 2.1. Requirements; 2.2. Metagenomic preparation; 2.3. Annotation; 2.4. CRISPR-Cas system identification and typing; 3. Visualize and characterize the repeat-spacer array; 3.1. Requirements; 3.2. Extract and visualize the repeat spacer array; 3.3. Determine CRISPR locus orientation; 4. crRNA guides; 4.1. Requirements; 4.2. tracrRNA identification
  • 4.3. crRNA boundaries and tracrRNA identification through RNASeq5. Predicting the PAM sequence; 5.1. Requirements; 5.2. Procedure; Acknowledgments; References; Chapter Two: Reconstitution and biochemical characterization of ribonucleoprotein complexes in Type I-E CRISPR-Cas systems; 1. Introduction; 1.1. In vivo assay for Cascade- and Cas3-mediated target plasmid loss; 1.2. Cloning, expression, and purification of the T. fusca Cascade complex; 2. Electrophoretic mobility shift assay; 2.1. Chemical probing
  • 2.2. Assemble TfuCascade/seed-bubble, TfuCascade/R-loop, and TfuCascade/R-loop/Cas3 complexes for structural analysis3. Conclusion; Acknowledgments; References; Further reading; Chapter Three: Sortase-mediated fluorescent labeling of CRISPR complexes; 1. Introduction; 2. Methods; 2.1. Overview; 2.2. Buffers; 2.2.1. Sortase purification buffers; 2.2.2. Cas1-Cas2 purification buffers; 2.2.3. Cascade purification buffers; 2.2.4. Cas3 purification buffers; 2.3. Sortase purification; 2.4. Purification of Tfu Cascade complexes and subunits for N-terminal sortase labeling
  • 2.4.1. Cas1-Cas2 purification and sortase labeling of Cas22.4.2. Cascade purification and sortase labeling of Cse1; 2.5. Purification of Tfu Cascade complexes and subunits for C-terminal sortase labeling; 2.5.1. Cas3 purification and sortase labeling; 2.6. Optimization of sortase-mediated fluorescent labeling; 3. Applications; 3.1. Single-molecule imaging of fluorescent Cas1-Cas2, Cascade, and Cas3 on DNA curtains; 4. Notes; Acknowledgments; Funding; References; Chapter Four: Fluorescence-based methods for measuring target interference by CRISPR-Cas systems; 1. Introduction
  • 2. Fluorescence-based strategies for measuring CRISPR interference2.1. Design and development of GFP-reporter plasmid pACYC-GFP; 2.2. Validation of GFP-based plasmid-loss assay; 3. Measurement of CRISPR interference in colonies and liquid culture; 3.1. Addition of CRISPR target to pACYC-GFP; 3.1.1. Equipment; 3.1.2. Buffers and reagents; 3.1.3. Procedure; 3.2. Detection of plasmid levels in bacterial colonies; 3.2.1. Equipment; 3.2.2. Buffers and reagents; 3.2.3. Procedure; 3.3. Measurement of CRISPR interference efficiency in liquid cultures; 3.3.1. Equipment; 3.3.2. Buffers and reagents
Control code
1083641420
Dimensions
unknown
Edition
First edition.
Extent
1 online resource
File format
unknown
Form of item
online
Isbn
9780128167618
Level of compression
unknown
Media category
computer
Media MARC source
rdamedia
Media type code
  • c
http://library.link/vocab/ext/overdrive/overdriveId
9780128167618
Quality assurance targets
not applicable
Reformatting quality
unknown
Sound
unknown sound
Specific material designation
remote
System control number
(OCoLC)1083641420
Label
CRISPR-Cas Enzymes, edited by Scoll Bailey
Publication
Antecedent source
unknown
Bibliography note
Includes bibliographical references at the end of each chapter and indexes
Carrier category
online resource
Carrier category code
  • cr
Carrier MARC source
rdacarrier
Color
multicolored
Content category
text
Content type code
  • txt
Content type MARC source
rdacontent
Contents
  • Front Cover; CRISPR-Cas Enzymes; Copyright; Contents; Contributors; Preface; Chapter One: Predicting and visualizing features of CRISPR-Cas systems; 1. Introduction; 2. Identify and characterize CRISPR-Cas system enzymes; 2.1. Requirements; 2.2. Metagenomic preparation; 2.3. Annotation; 2.4. CRISPR-Cas system identification and typing; 3. Visualize and characterize the repeat-spacer array; 3.1. Requirements; 3.2. Extract and visualize the repeat spacer array; 3.3. Determine CRISPR locus orientation; 4. crRNA guides; 4.1. Requirements; 4.2. tracrRNA identification
  • 4.3. crRNA boundaries and tracrRNA identification through RNASeq5. Predicting the PAM sequence; 5.1. Requirements; 5.2. Procedure; Acknowledgments; References; Chapter Two: Reconstitution and biochemical characterization of ribonucleoprotein complexes in Type I-E CRISPR-Cas systems; 1. Introduction; 1.1. In vivo assay for Cascade- and Cas3-mediated target plasmid loss; 1.2. Cloning, expression, and purification of the T. fusca Cascade complex; 2. Electrophoretic mobility shift assay; 2.1. Chemical probing
  • 2.2. Assemble TfuCascade/seed-bubble, TfuCascade/R-loop, and TfuCascade/R-loop/Cas3 complexes for structural analysis3. Conclusion; Acknowledgments; References; Further reading; Chapter Three: Sortase-mediated fluorescent labeling of CRISPR complexes; 1. Introduction; 2. Methods; 2.1. Overview; 2.2. Buffers; 2.2.1. Sortase purification buffers; 2.2.2. Cas1-Cas2 purification buffers; 2.2.3. Cascade purification buffers; 2.2.4. Cas3 purification buffers; 2.3. Sortase purification; 2.4. Purification of Tfu Cascade complexes and subunits for N-terminal sortase labeling
  • 2.4.1. Cas1-Cas2 purification and sortase labeling of Cas22.4.2. Cascade purification and sortase labeling of Cse1; 2.5. Purification of Tfu Cascade complexes and subunits for C-terminal sortase labeling; 2.5.1. Cas3 purification and sortase labeling; 2.6. Optimization of sortase-mediated fluorescent labeling; 3. Applications; 3.1. Single-molecule imaging of fluorescent Cas1-Cas2, Cascade, and Cas3 on DNA curtains; 4. Notes; Acknowledgments; Funding; References; Chapter Four: Fluorescence-based methods for measuring target interference by CRISPR-Cas systems; 1. Introduction
  • 2. Fluorescence-based strategies for measuring CRISPR interference2.1. Design and development of GFP-reporter plasmid pACYC-GFP; 2.2. Validation of GFP-based plasmid-loss assay; 3. Measurement of CRISPR interference in colonies and liquid culture; 3.1. Addition of CRISPR target to pACYC-GFP; 3.1.1. Equipment; 3.1.2. Buffers and reagents; 3.1.3. Procedure; 3.2. Detection of plasmid levels in bacterial colonies; 3.2.1. Equipment; 3.2.2. Buffers and reagents; 3.2.3. Procedure; 3.3. Measurement of CRISPR interference efficiency in liquid cultures; 3.3.1. Equipment; 3.3.2. Buffers and reagents
Control code
1083641420
Dimensions
unknown
Edition
First edition.
Extent
1 online resource
File format
unknown
Form of item
online
Isbn
9780128167618
Level of compression
unknown
Media category
computer
Media MARC source
rdamedia
Media type code
  • c
http://library.link/vocab/ext/overdrive/overdriveId
9780128167618
Quality assurance targets
not applicable
Reformatting quality
unknown
Sound
unknown sound
Specific material designation
remote
System control number
(OCoLC)1083641420

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