This second volume provides new and updated methods detailing advancements in CRISPR-Cas technical protocols. Chapters guide readers through protocols on prime editing, base editing, multiplex editing, editing in cell-free extract, in silico analysis of gRNA secondary structure and CRISPR-diagnosis.
Authoritative and cutting-edge, CRISPR-Cas Methods, Volume 2 aims to serves as a laboratory manual providing scientists with a holistic view of CRISPR-Cas methodologies and its practical application for the editing of crop plants, cell lines, nematode and microorganism.
The chapter "CRISPR/Cas9-mediated gene editing in human induced pluripotent stem cells" is available open access under a Creative Commons Attribution 4. 0 International License via link. springer. com.
Inhaltsverzeichnis
A Complete Methodology for the Instruction of CRISPR-based Gene Editing Using a Simplified Cell-free Extract System with Genetic Readout in Bacteria. -
In silico
Analysis of gRNA Secondary Structure to Predict Its Efficacy for Plant Genome Editing. - Efficient Multiplexed CRISPR-Cas12a Genome Editing in Plants. - CRISPR-dCas9 Based Targeted Manipulation of DNA Methylation in Plants. - Rapid Assembly of Multiplex Natural CRISPR Arrays. - Assembly and Assessment of Prime Editing Systems for Precise Genome Editing in Plants. - Designing, Performing and Analysing CRISPR-Cas9-mediated Genome Editing Experiments in Leguminous Plant. - Generation of Knockout and Fragment Deletion Mutants in Soybean by CRISPR-Cas9. - Targeted Base Editing in Soybean using a CRISPR-Cas9 Cytidine Deaminase Fusion. - Genome Editing in Potato Using a Hairy Root Transformation System. - Efficient CRISPR-Cas9-Mediated Genome Editing in Tomato. - CRISPR-Cas9-Mediated Genome Editing in Rice: A Systematic Protocol for Single and Multi-Target Vector Construction. - Generating Clonal Seeds from Hybrid Rice with CRISPR-Cas9. - CRISPR-Cas12a-based DNA Detection for Fast Pathogen Diagnosis and GMO Test in Plants. - CRISPR-Cas9-mediated Gene Editing in Human Induced Pluripotent Stem Cells. - Cell-Type-Specific CRISPR-Cas9 System with miRNAs. - CRISPR-Cas9 gene Editing in Mammalian Cells Using LentiCRISPRv2/LentiGuide-puro Vectors. - An Approach to Proximity Ligation by T4 RNA Ligase to Screen sRNA that Regulate CRISPR-Cas systems. - Genetic Engineering of a Phage-based Delivery System for Endogenous III-A CRISPR-Cas System against
Mycobacterium tuberculosis. -
CRISPR-Cas9-Mediated Genome Editing in
Escherichia coli
Bacteriophages. - CRISPR-Cas Genome Editing in the Cellulolytic Bacterium
Clostridium thermocellum. -
Gene Targeting in
Caenorhabditis elegans
Using a Combination of Multiple sgRNAs and a Homologous Recombination-mediated Repair. - Design of Repair Templates for CRISPR-Cas9-triggered Homologous Recombination in
Caenorhabditis elegans.
