1. A Complete Methodology for the Instruction of CRISPR-based Gene Editing Using a Simplified Cell-free Extract System with Genetic Readout in Bacteria
Kristen M. Pisarcik, Brett M. Sansbury and Eric B. Kmiec
2. In silico Analysis of gRNA Secondary Structure to Predict Its Efficacy
for Plant Genome Editing
Md Mahmudul Hassan, Abul Kashem Chowdhury and Tofazzal Islam
3. Efficient Multiplexed CRISPR-Cas12a Genome Editing in Plants
Yingxiao Zhang and Yiping Qi
4. CRISPR-dCas9 Based Targeted Manipulation of DNA Methylation in Plants
Basudev Ghoshala and Jason Gardinera
5. Rapid Assembly of Multiplex Natural CRISPR Arrays
Robert M. Cooper and Jeff Hasty
6. Assembly and Assessment of Prime Editing Systems for Precise Genome Editing in Plants
Simon Sretenovic, Changtian Pan, and Yiping Qi
7. Designing, Performing and Analysing CRISPR-Cas9-mediated Genome Editing Experiments in Leguminous Plant
Dipak Gayen and Subhasis Karmakar
8. Generation of Knockout and Fragment Deletion Mutants in Soybean by CRISPR-Cas9
Li Chen, Yupeng Cai and Wensheng Hou
9. Targeted Base Editing in Soybean using a CRISPR-Cas9 Cytidine Deaminase Fusion
Li Chen, Yupeng Cai and Wensheng Hou
10. Genome Editing in Potato Using a Hairy Root Transformation System
Nathaniel M. Butler and Jiming Jiang
11. Efficient CRISPR-Cas9-Mediated Genome Editing in Tomato
Viswanathan Satheesh, Jinkai Li and Mingguang Lei
12. CRISPR-Cas9-Mediated Genome Editing in Rice: A Systematic Protocol for Single and Multi-Target Vector Construction
Hirendra Nath Barman
13. Generating Clonal Seeds from Hybrid Rice with CRISPR-Cas9
Chaolei Liu, Chun Wang and Kejian Wang
14. CRISPR-Cas12a-based DNA Detection for Fast Pathogen Diagnosis and GMO Test in Plants
Yun-Mu Zhang, Yinong Yang and Kabin Xie
15. CRISPR-Cas9-mediated Gene Editing in Human Induced Pluripotent Stem Cells
Stijn L.M. in ’t Groen, Mike Broeders, and W.W.M. Pim Pijnappel
16. Cell-Type-Specific CRISPR-Cas9 System with miRNAs
Moe Hirosawa and Hirohide Saito
17. CRISPR-Cas9 gene Editing in Mammalian Cells Using LentiCRISPRv2/LentiGuide-puro Vectors
Zubaida Sa’id Ameen, Ece Çakiroglu, Serif Senturk, Abdullahi Umar Ibrahhim and Mehmet Ozsoz
18. An Approach to Proximity Ligation by T4 RNA Ligase to Screen sRNA that Regulate CRISPR-Cas systems
Ping Lin, Qinqin Pu, and Min Wu
19. Genetic Engineering of a Phage-based Delivery System for Endogenous III-A CRISPR-Cas System against Mycobacterium tuberculosis
Li Zheng Jiang, Junwei Wei, Nan Peng and Yingjun Li
20. CRISPR-Cas9-Mediated Genome Editing in Escherichia coli Bacteriophages
Yibao Chen, Xiangmin Li, Shuang Wang, Ping Qian, Yingjun Li
21. CRISPR-Cas Genome Editing in the Cellulolytic Bacterium Clostridium thermocellum
Jiang Sean Stettner and Carrie Eckert
22. Gene Targeting in Caenorhabditis elegans Using a Combination of Multiple sgRNAs and a Homologous Recombination-mediated Repair
Jose Luis Tellez-Arreola
23. Design of Repair Templates for CRISPR-Cas9-triggered Homologous Recombination in Caenorhabditis elegans
Li Hyun-Min Kim and Xiaojuan Zhang
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.”