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E-grāmata: CRISPR Guide RNA Design: Methods and Protocols

  • Formāts: PDF+DRM
  • Sērija : Methods in Molecular Biology 2162
  • Izdošanas datums: 14-Sep-2020
  • Izdevniecība: Springer-Verlag New York Inc.
  • Valoda: eng
  • ISBN-13: 9781071606872
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  • Formāts: PDF+DRM
  • Sērija : Methods in Molecular Biology 2162
  • Izdošanas datums: 14-Sep-2020
  • Izdevniecība: Springer-Verlag New York Inc.
  • Valoda: eng
  • ISBN-13: 9781071606872
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This detailed volume focuses on the CRISPR-associated guide RNA and how it can be designed, modified, and validated for a broad repertoire of purposes. Beginning with a section on computational design of target-specific guide RNAs, the book continues by covering chemical modifications to alter guide RNA stability, specificity, and efficiency, as well as to create inducible guide RNAs, append additional functional domains, and express guide RNAs in a conditional manner. It concludes with methods for measuring off-target guide RNA activity. 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 protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and essential, CRISPR Guide RNA Design: Methods and Protocols provides a comprehensive pipeline for guide RNA design and aims to be an invaluable resource in applying this powerful technology to basic research and therapeutic applications.
Preface v
Contributors ix
PART I IN SILICO DESIGN AND OPTIMIZATION OF GUIDE RNA SEQUENCES
1 Cloud-Based Design Of Short Guide Rna (Sgrna) Libraries For Crispr Experiments
3(20)
Florian Heigwer
Michael Boutros
2 Web-Based Crispr Toolkits: Cas-Offinder, Cas-Designer, And Cas-Analyzer
23(14)
Gue-Ho Hwang
Jin-Soo Kim
Sangsu Bae
PART II CHEMICALLY-MODIFIED GUIDE RNAS
3 Chemical Modification Of Guide Rnas For Improved Crispr Activity In Cd34+ Human Hematopoietic Stem And Progenitor Cells
37(12)
Jenny Shapiro
Adi Tovin
Ortal Iancu
Daniel Allen
Ayal Hendel
4 Gene Disruption Using Chemically Modified Crispr-Cpfl Rna
49(12)
Moira A. Mcmahon
Meghdad Rahdar
5 "Split-And-Click" Sgrna
61(18)
Lapatrada Taemaitree
Arun Shivalingam
Afaf H. El-Sagheer
Tom Brown
6 Chimeric Dna-Rna Guide Rna Designs
79(10)
Shuhan Lu
Ying Zhang
Hao Tin
PART III EXPANDING THE CRISPR TOOLBOX
7 Harnessing Trna For Processing Ability And Promoter Activity
89(26)
David J. H. F. Knapp
Tudor A. Fulga
8 Targeting Noncoding Rna Domains To Genomic Loci With Crispr-Display: Guidelines For Designing, Building, And Testing Sgrna-Ncrna Expression Constructs
115(38)
David M. Shechner
9 Controlling The Activity Of Crispr Transcriptional Regulators With Inducible Sgrnas
153(32)
Quentin R. V. Ferry
Tudor A. Fulga
10 Gene Manipulation Using Fusion Guide Rnas For Cas9 And Cas 12A
185(12)
Ha Rim Shin
Jiyeon Kweon
Yongsub Kim
PART IV CHARACTERIZATION OF CRISPR EFFICACY AND SPECIFICITY
11 Methods For Measuring Crispr/Cas9 Dna Cleavage In Cells
197(18)
Christopher R. Cromwell
Juan Jovel
Basil P. Hubbard
12 In Vitro Assays For Comparing The Specificity Of First- And Next-Generation Crispr/Cas9 Systems
215(18)
Christopher R. Cromwell
Basil P. Hubbard
13 Profiling Genome-Wide Specificity Of Crispr-Cas9 Using Digenome-Seq
233(10)
Daesik Kim
Jin-Soo Kim
14 Detection Of Crispr/Cas9-Generated Off-Target Effect By Integration-Defective Lentiviral Vector
243(18)
Xiaoling Wang
Youjun Wu
Jiing-Kuan Tee
15 Genome-Wide Crispr Off-Target Dna Break Detection By The Bliss Method
261(22)
Roberto Ballarino
Britta A. M. Bouwman
Nicola Crosetto
Index 283