فهرست مطالب:

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
2 Web-Based CRISPR Toolkits: Cas-OFFinder, Cas-Designer, and Cas-Analyzer . . . . . 23
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
4 Gene Disruption Using Chemically Modified CRISPR-Cpf1 RNA . . . . . . . . . . . . 49
5 “Split-and-Click” sgRNA . . . . . . .  . . . . . 61
6 Chimeric DNA–RNA Guide RNA Designs. . . . . . . . . . . . . 79
PART III EXPANDING THE CRISPR TOOLBOX
7 Harnessing tRNA for Processing Ability and Promoter Activity. . . . . . . . . . . . . . . 89
8 Targeting Noncoding RNA Domains to Genomic Loci with CRISPR-Display: Guidelines for Designing, Building, and Testing sgRNA–ncRNA Expression Constructs . . . . . . . 115
9 Controlling the Activity of CRISPR Transcriptional Regulators with Inducible sgRNAs . . . . 153
10 Gene Manipulation Using Fusion Guide RNAs for Cas9 and Cas12a. . . . . . . . . . 185
PART IV CHARACTERIZATION OF CRISPR EFFICACY AND SPECIFICITY
11 Methods for Measuring CRISPR/Cas9 DNA Cleavage in Cells . . . . . . . . . . . . . . 197
12 In Vitro Assays for Comparing the Specificity of First- and Next-Generation CRISPR/Cas9 Systems. . . . 215
13 Profiling Genome-Wide Specificity of CRISPR-Cas9 Using Digenome-Seq . . . . 233
14 Detection of CRISPR/Cas9-Generated Off-Target Effect by Integration-Defective Lentiviral Vector . . . . 243
15 Genome-Wide CRISPR Off-Target DNA Break Detection by the BLISS Method . . . 261
Index . . . . . . . . 283

مشخصات فایل