This second edition provides new and updated protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Genome Editing in Animals: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.
Construction and Evaluation of Zinc Finger Nucleases.- Updated overview of TALEN construction systems.- CRISPR/Cas9.- SNPD-CRISPR: Single Nucleotide Polymorphism-Distinguishable Repression or Enhancement of a Target Gene Expression by CRISPR System.- Utilizing large functional and population genomics resources for CRISPR/Cas perturbation experiment design.- Generation of genome-edited mice by cytoplasmic injection of CRISPR-Cas9 RNA.- Gene targeting in mouse embryonic stem cells via CRISPR/Cas9 ribonucleoprotein (RNP) mediated genome editing.- Generation of Knock-in Mouse by Genome Editing.- Introduction of genetic mutations into mice by Base Editor and Target-AID.- Genome editing in mouse and rat by electroporation.- Generation of floxed mice by sequential electroporation.- Efficient detection of flox mice using in vitro Cre recombination.- VCre/VloxP and SCre/SloxP as reliable site-specific recombination systems for genome engineering.- Efficient CRISPR/Cas9-Assisted Knockin ofLarge DNA Donors by Pronuclear Microinjection During S-phase in Mouse Zygotes.- Genome editing of murine liver hepatocytes by AAV vector-mediated expression of Cas9 in vivo.- Non-Viral Ex Vivo Genome-Editing in Mouse Bona Fide Hematopoietic Stem Cells with CRISPR/Cas9.- Genome editing of rat.- A simple and efficient method for generating KO rats using in vitro fertilized oocytes.- Editing the Genome of the Golden Hamster (Mesocricetus auratus).- Gene targeting in rabbits: single-step generation of knockout rabbits by microinjection of CRISPR/Cas9 plasmids.- Genome Editing of Pig.- GEEP method: an optimized electroporation-mediated gene-editing approach for establishment of knockout pig lines.- Genome editing mediated by primordial germ cell in chicken.- CRISPR-Cas9-mediated genome modifications in zebrafish.- Genome Editing of Medaka. -CRISPR-Cas9 based functional analysis in amphibians; Xenopus laevis, Xenopus tropicalis, and Pleurodeles waltl.- Genome Editing of Silkworms.- Improved genome editing in the ascidian Ciona with CRISPR/Cas9 and TALEN.- Genome editing of C. elegans.
Inhaltsverzeichnis
1. Construction and Evaluation of Zinc Finger Nucleases
Hiroshi Ochiai and Takashi Yamamoto
2. Updated overview of TALEN construction systems
Tetsushi Sakuma and Takashi Yamamoto
3. CRISPR/Cas9
Izuho Hatada, Sumiyo Morita, and Takuro Horii
4 SNPD-CRISPR: Single Nucleotide Polymorphism-Distinguishable Repression or Enhancement of a Target Gene Expression by CRISPR System
Shohei Maruyama, Takashi Kusakabe, Xinyi Zou, Yoshiaki Kobayashi, Yoshimasa Asano, Qingbo S. Wang, and Kumiko Ui-Tei
5. Utilizing large functional and population genomics resources for CRISPR/Cas perturbation experiment design
Qingbo S. Wang, Kumiko Ui-Tei
6. Generation of genome-edited mice by cytoplasmic injection of CRISPR-Cas9 RNA
Takuro Horii and Izuho Hatada
7. Gene targeting in mouse embryonic stem cells via CRISPR/Cas9 ribonucleoprotein (RNP) mediated genome editing.
Manabu Ozawa, Chihiro Emori, and Masahito Ikawa
8. Generation of Knock-in Mouse by Genome Editing
Wataru Fujii
9. Introduction of genetic mutations into mice by Base Editor and Target-AID
Hiroki Sasaguri
10. Genome editing in mouse and rat by electroporation
Takehito Kaneko
11. Generation of floxed mice by sequential electroporation
Takuro Horii, Ryosuke Kobayashi, and Izuho Hatada
12. Efficient detection of flox mice using in vitro Cre recombination
Ryosuke Kobayashi, Takuro Horii, and Izuho Hatada
13. VCre/VloxP and SCre/SloxP as reliable site-specific recombination systems for genome engineering
Manabu Nakayama
14. Efficient CRISPR/Cas9-Assisted Knockin of Large DNA Donors by Pronuclear Microinjection During S-phase in Mouse Zygotes
Takaya Abe, Ken-ichi Inoue, and Hiroshi Kiyonari
15. Genome editing of murine liver hepatocytes by AAV vector-mediated expression of Cas9 in vivo
Yuji Kashiwakura and Tsukasa Ohmori
16. Non-Viral Ex Vivo Genome-Editing in Mouse Bona Fide Hematopoietic Stem Cells with CRISPR/Cas9
Hiromasa Hara1, Natsagdorj Munkh-Erdene, Suvd Byambaa, and Yutaka Hanazono
17. Genome editing of rat
Takehito Kaneko
18. A simple and efficient method for generating KO rats using in vitro fertilized oocytes
Kohtaro Morita, Arata Honda, and Masahide Asano
19. Editing the Genome of the Golden Hamster (Mesocricetus auratus)
Michiko Hirose, Toshiko Tomishima, and Atsuo Ogura
20. Gene targeting in rabbits: single-step generation of knockout rabbits by microinjection of CRISPR/Cas9 plasmids
Arata Honda
21. Genome Editing of Pig
Masahito Watanabe and Hiroshi Nagashima
22. GEEP method: an optimized electroporation-mediated gene-editing approach for establishment of knockout pig lines
Fuminori Tanihara, Maki Hirata, and Takeshige Otoi
23. Genome editing mediated by primordial germ cell in chicken
Jae Yong Han and Hong Jo Lee
24. CRISPR¿Cas9-mediated genome modifications in zebrafish
Yusuke Kamachi and Atsuo Kawahara
25. Genome Editing of Medaka
Rie Hara, Satoshi Ansai, and Masato Kinoshita
26. CRISPR-Cas9 based functional analysis in amphibians; Xenopus laevis, Xenopus tropicalis, and Pleurodeles waltl
Miyuki Suzuki, Midori Iida, Toshinori Hayashi, and Ken-ichi T Suzuki
27. Genome Editing of Silkworms
Takuya Tsubota, Hiroki Sakai, and Hideki Sezutsu
28. Improved genome editing in the ascidian Ciona with CRISPR/Cas9 and TALEN
Yasunori Sasakura and Takeo Horie
29. Genome editing of C. elegans
Takuma Sugi
Klappentext
This second edition provides new and updated protocols that can be used for generation of knockout animals. Chapters guide the reader through basic protocols for three genome editing technologies, target design tools, and specific protocols for each animal. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Genome Editing in Animals: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.
