CRISPR-Cas Methods

1. Front Matter

   Pages i-xiv

   PDF

2. Wide Horizons of CRISPR-Cas-Derived Technologies for Basic Biology, Agriculture, and Medicine

   • Kutubuddin A. Molla, Subhasis Karmakar, M. Tofazzal Islam

   Pages 1-23

3. Efficient Homologous Recombination-Mediated in Planta Gene Targeting by Egg-Cell-Specific Expression of Staphylococcus aureus Cas9 from Arabidopsis

   • Felix Wolter, Teng-Kuei Huang, Holger Puchta

   Pages 25-34

4. Rice Gene Knockout or Downregulation through CRISPR-Cas9

   • Yvonne Ludwig, Inez H. Slamet-Loedin

   Pages 35-50

5. CRISPR-Cas-Mediated Single Base Editing at More than One Locus in Rice Genome

   • Kutubuddin A. Molla, Yinong Yang

   Pages 51-62

6. CRISPR-Cas9-Mediated Genome Editing of the Model Grass Species Brachypodium distachyon

   • Deborah Petrik

   Pages 63-86

7. CRISPR-Cas9-Mediated Gene Editing of the Plant Pathogenic Oomycete Phytophthora palmivora

   • Miaoying Tian, Natasha Navet, Dongliang Wu

   Pages 87-98

8. An Effective CRISPR-Cas9 Technology for Efficiently Isolating Transgene-Free Mutants in Arabidopsis, Brassica napus, Strawberry, and Soybean

   • Cheng Dai, Hong Yang, Ting Tang, Chaozhi Ma, Chunying Kang

   Pages 99-115

9. An Optimized RNA-Guided Cas9 System for Efficient Simplex and Multiplex Genome Editing in Barley (Hordeum vulgare L.)

   • Sebastian Gasparis, Mateusz Przyborowski

   Pages 117-142

10. Genome Editing of Mammalian Cells Using CRISPR-Cas: From In Silico Designing to In-Culture Validation

    • Borhan Uddin, Patrick Partscht, Taslima Nahar

    Pages 143-162

11. Cloning-Free (DNA-Free) CRISPR-Cas9-Mediated Gene Editing in Human Liver Cell Line and Its Detection

    • Magdalena Śmiech, Paweł Leszczyński, Effi Haque, Hiroaki Taniguchi

    Pages 163-171

12. A Procedure to Design Guide RNA, Assemble Fragments, and Detect Mutation for Genome Editing in Flax

    • Dinesh Adhikary, Isadora Louise Alves da Costa Ribeiro Quintans, Pankaj K. Bhowmik

    Pages 173-190

13. Delivery of Cas9/sgRNA RNP into Wheat Microspores Using Synthetic CPP for Genome Editing and Gene Expression Modulation

    • Andriy Bilichak, Justin Luu, Fengying Jiang, John Laurie

    Pages 191-202

14. CRISPR-Cas9-Mediated Gene Editing in Wheat: A Step-by-Step Protocol

    • Pankaj K. Bhowmik, M. Tofazzal Islam

    Pages 203-222

15. CRISPR-Cas9-Based Genome Editing of Banana

    • Leena Tripathi, Valentine Otang Ntui, Jaindra Nath Tripathi

    Pages 223-236

16. Homology-Directed Transgene-Free Gene Editing in Chlamydomonas reinhardtii

    • Aron Ferenczi, Attila Molnar

    Pages 237-252

17. Correction to: A Procedure to Design Guide RNA, Assemble Fragments, and Detect Mutation for Genome Editing in Flax

    • Dinesh Adhikary, Isadora Louise Alves da Costa Ribeiro Quintans, Pankaj K. Bhowmik

    Pages C1-C1

18. Back Matter

    Pages 253-278

    PDF

This volume details the fundamentals of the CRISPR-Cas system, and its protocols illustrate advances in CRISPR-Cas techniques for efficient genome editing. Introductory chapters provide a wide horizon of CRISPR/Cas-based methods and applications. Additional chapters guide readers through HDR-mediated editing, sgRNA design, the step-by-step procedure of multiplex adenine base editing experiments in rice, generating mutants for rice, wheat, Brachypodium, Barley, Flax, and Phytophthora, visual screening of mutants, gene deletion (knock-out), tagging (knock-in) in mammalian cells, the cloning-free (DNA-free) technique, cell-penetrating peptides, generating a genome-edited banana, and nuclear genome editing of Chlamydomonas employing CRISPR-Cpf1 combined with a single-stranded DNA (ssODN) repair template. 


Authoritative and cutting-edge, CRISPR-Cas Methods aims to assist researchers who are new to the field and are aiming to learn how best to adopt this technology for a particular organism.

• Staphylococcus aureus Cas9
• Brachypodium distachyon
• P. palmivora mutants
• deletion (knock-out)
• tagging (knock-in)

• Institute of Biotechnology and Genetic Engineering (IBGE), Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh

  M. Tofazzal Islam

• Department of Aquatic and Crop Resource Development, National Research Council of Canada, SASKATOON, Canada

  Pankaj K. Bhowmik

• Crop Improvement Division, ICAR-National Rice Research Institute, Cuttack, India

  Kutubuddin A. Molla

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