CRISPR Handbook : Applications, Tools, and Resources / Zhi-Yan Du.

Format:

Book

Author/Creator:

Du, Zhi-Yan, author.

Language:

English

Subjects (All):

CRISPR (Genetics).

Physical Description:

1 online resource (159 pages)

Edition:

First edition.

Other Title:

Applications, Tools, and Resources

Place of Publication:

London, England : Academic Press, 2024.

Summary:

CRISPR Handbook: Applications, Tools and Resources presents a detailed introduction to the CRISPR technology. This handbook summarizes the current CRISPR applications in diverse organisms, shares useful tools for starting a CRISPR experiment, and provides information on software and services for CRISPR technology. Written by an expert in the field of biotechnology and molecular biology of diverse experiment models, including plants, microalgae, and fungi. This handbook will serve as a guide for those who are interested in learning about CRISPR technologies and their applications in research and industry.

Contents:

Front Cover

CRISPR Handbook

Copyright Page

Contents

Acknowledgments

1 What is CRISPR?

1.1 Introduction to genome-editing technology

1.1.1 The history of genome-editing technology

1.1.2 Overview of CRISPR technology

1.2 The history of CRISPR

1.3 The components of CRISPR and how it works

1.3.1 sgRNA

1.3.2 PAM sequence

1.3.3 Cas protein

1.3.4 Classifications of CRISPR/Cas systems

1.3.5 Delivery methods

1.3.5.1 Agrobacterium-mediated delivery

1.3.5.2 Particle bombardment

1.3.5.3 Single-cell microinjection

1.3.5.4 PEG-mediated delivery

1.3.5.5 Electroporation

1.3.5.6 Viral vectors

1.3.5.7 Nanoparticles

1.4 Challenges and ethical impacts of CRISPR technology

1.4.1 Off-target effects of CRISPR and delivery

1.4.2 Ethics and societal concerns

1.4.3 Safety concerns and regulation

References

2 Applications of CRISPR technology

2.1 CRISPR/Cas-mediated gene repression and activation

2.1.1 CRISPR interference and activation

2.1.2 Epigenetic editing with CRISPR

2.2 Live imaging

2.2.1 Overview of CRISPR/Cas for live imaging

2.2.2 Challenges for CRISPR-based live imaging

2.3 Therapeutic applications

2.3.1 Applications of CRISPR for genetic disease therapies

2.3.2 CRISPR against viral infection

2.3.3 Cancer applications

2.3.3.1 Cancer detection and monitoring

2.3.3.2 Cancer treatment

2.3.3.3 Cancer research

2.4 CRISPR diagnostics

2.4.1 SHERLOCK and HUDSON systems

2.4.2 DETECTR system

2.4.3 Challenges of CRISPR-Dx

2.5 Agriculture

2.5.1 Disease resistance

2.5.2 Crop yield traits

2.5.3 Nutrition and palatability

2.5.4 Transgene-free gene editing

2.5.5 De novo domestication

2.5.6 Livestock industry

2.6 Synthetic biology

2.7 Other applications

2.7.1 High-throughput screening of molecular targets.

2.7.2 Pest control

2.7.3 Bioremediation

2.7.4 Organ transplantation

2.7.5 Lab-grown meat

3 CRISPR applications for various organisms

3.1 CRISPR applications for prokaryotes

3.2 CRISPR applications for eukaryotes

3.2.1 Fungi

3.2.2 Plantae

3.2.3 Animalia

4 Tools and resources for CRISPR technology

4.1 Genomes and gene sequences

4.2 gRNA design and synthesis resources

4.3 Sequence software

4.4 Evaluation and screening

4.5 Data analysis

4.6 Services

5 Protocols for CRISPR technology

5.1 Saccharomyces cerevisiae (yeast)

5.1.1 Methods

5.1.1.1 Preparation of Cas12a plasmids

5.1.1.2 Preparation of the single crRNA array expression cassette

5.1.1.3 Preparation of promoter-ORF-terminator donor DNA constructs

5.1.1.4 Preparation of integration flank DNA sequences containing connector sequences

5.1.1.5 S. cerevisiae transformation

5.1.1.6 Efficiency evaluation of genome editing

5.1.1.7 Confirmation of integration of donor DNA at the intended loci

5.1.2 List/s of materials

5.2 Zea mays (maize)

5.2.1 Methods

5.2.1.1 Growth media preparation

5.2.1.2 Growing donor maize plants and harvesting immature ears

5.2.1.3 CRISPR/Cas9 and Cas12a vector construction

5.2.1.4 Preparing Agrobacterium suspension culture for infection

5.2.1.5 Embryo dissection, infection, and co-cultivation

5.2.1.6 Heat treatment, transformant selection, and regeneration

5.2.1.7 Greenhouse and T1 seeds

5.2.1.8 On- and off-target mutagenesis analysis

5.2.2 List/s of materials

5.3 Mus musculus (mouse)

5.3.1 Methods

5.3.1.1 gRNA design for the CRISPR-concatemer vector

5.3.1.2 Cloning of gRNAs into the CRISPR-concatemer vector

5.3.1.3 Transfection of intestinal organoids by electroporation.

5.3.1.4 Growth factor withdrawal (phenotype of mutants)

5.3.2 List/s of materials

Index

Back Cover.

Notes:

Includes bibliographical references and index.

Description based on publisher supplied metadata and other sources.

ISBN:

9780443238666

0443238669

OCLC:

1472986384