Crop Genome Editing Using CRISPR/Cas9: Theory and Practice is a highly useful reference for implementing genome engineering technologies, particularly CRISPR related projects in agricultural crops and other plants. This book provides an introduction to CRISPR's basic science and applied aspects, along with detailed protocols. It presents a detailed workflow, beginning with genome sequence retrieval and then mutation analysis in genome edited events using sequencing tools. The book helps those in the field methodically plan, design and conduct experiments. This practical guide will dramatically help researchers in accelerating conventional plant breeding programs.
- Offers a detailed review of literature on genome editing tools, with special emphasis paid to CRISPR/Cas9 and its advancements
- Contains step-by-step guidelines for single guide RNA design, CRISPR vector construction, protoplast transformation, mutation analysis and Agrobacterium-based regeneration of mutant plants
- Includes detailed troubleshooting tips during various steps
Table of Contents
1. Introduction to genome editing 2. Crop improvement using CRISPR/Cas9 3. Designing of sgRNA 4. Tools and Resources for the construction of CRISPR vectors 5. Cloning of CRISPR constructs 6. Protoplast Isolation from plants 7. Delivery of CRISPR constructs into protoplasts and mutation analysis 8. Agrobacterium mediated transformation of CRISPR constructs for the generation of stable mutant plants 9. Recent Advancements in CRISPR/Cas9 genome editing in plants
