CRISPR Gene Editing: Methods and Protocols

by Luo, Yonglun

Type Book

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Title Statements

Full title :CRISPR Gene Editing : Methods and Protocols.

Summary

This detailed volume guides readers through strategic planning and user-friendly guidelines in order to select the most suitable CRISPR-Cas system and target sites with high activity and specificity. Methods covering CRISPR gRNA design, CRISPR delivery, CRISPR activity quantification (indel quantification), and examples of applying CRISPR gene editing in human pluripotent stem cells, primary cells, gene therapy, and genetic screening are included. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and invaluable, CRISPR Gene Editing: Methods and Protocols will assist undergraduates, graduates, and researchers with detailed guidelines and methods for the vitally important CRISPR gene editing field. Chapter 3 is available open access under a CC BY 4.0 license via link.springer.com.

Edition Statements

Publication Date:2019.

Publication Statements

Publisher(s):Springer
Place of Publication:New York, NY
Language:eng
Publication Date:2019.

Notes

Chapter 5: Functional Evaluation of CRISPR Activity by the Dual-Fluorescent Surrogate System: C-Check

The Physical Object

Description:1 online resource (356 pages)

Edition Identifiers

ISBN:9781493991709, 1493991701
DOI:10.1007/978-1-4939-9170-9.
oclc:on1122920718
LC Classification:RB155-155.8
Dewey Decimal:611.01816

Intro; Preface; Contents; Contributors; Part I: Methods for CRISPR-gRNA Design and Quantification of Activity; Chapter 1: CRISPR-gRNA Design; 1 Introduction; 2 Nuclease Types; 3 gRNA Efficiency; 4 gRNA Off Target; 5 gRNA Design; 6 gRNA Expression, Construction, and Delivery; 7 Design of Repair Template (Only for HDR Applications); 8 Functional Validation of gRNA; 9 On-Target Efficiency; 10 Off-Target Analysis; 11 Conclusion; References; Chapter 2: Tracking CRISPRś Footprints; 1 The Important Role of Endogenous DSB Repair Machineries in CRISPR Gene Editing

2 CRISPRś Footprint Detection by Mismatch-Specific Endonuclease Assay (MSE)3 CRISPRś Footprint Detection by TIDE and TIDER Assays; 4 CRISPRś Footprint Detection by IDAA Assay; 5 CRISPRś Footprint Detection by High Resolution Melting Curve; 6 CRISPRś Footprint Detection by Drop-Off Assay; 7 CRISPRś Footprint Detection by Surrogate Reporter Systems; 8 CRISPRś Footprint Detection by Targeted Amplicon Next-Generation Sequencing (TA-NGS); 9 Genome-Scale Detection of CRISPRś Footprints; 10 CRISPR Indels Are Not Random; 11 Concluding Remarks; References

Chapter 3: Rapid Quantitative Evaluation of CRISPR Genome Editing by TIDE and TIDE…