1. Microinjection and Micromanipulation: A Historical Perspective
Wenhao Xu
2. Production of Transgenic Mice by Pronuclear Microinjection
Xin-an Pu, Anthony P. Young, and H. Michael Kubisch
3. Transgene Recombineering in Bacterial Artificial Chromosomes
Michael G. Zeidler and Thomas L. Saunders
4. Using TARGATTTM Technology to Generate Site-Specific Transgenic Mice
Ruby Yanru Chen-Tsai
5. Generating Genetically Engineered Mice Using a Spermatogonial Stem Cell-Mediated Method
Takuya Sato and Takehiko Ogawa
6. Chimeric Mouse Generation by ES Cell Blastocyst Microinjection and Uterine Transfer
Yubin Du, Wen Xie, Fan Zhang, and Chengyu Liu
7. Creating Knock-In Alleles in Mouse Embryonic Stem Cells by CRISPR/Cas9-Mediated Homologous Recombination without Drug Selection
Pinghu Liu, Yan Li, Jingqi Lei, and Lijin Dong
8. Using CRISPR/Cas9 for Gene Knockout in Immunodeficient NSG Mice
Yubin Du, Wen Xie, Fan Zhang, Uimook Choi, Chengyu Liu, and Colin L. Sweeney
9. Generation of CRISPR-Edited Rodents Using a Piezo-Driven Zygote Injection Technique
Melissa A. Scott and Yueh-Chiang Hu
10. Delivery of CRISPR-Cas9 into Mouse Zygotes by Electroporation
Wenning Qin and Haoyi Wang
11. Generation of Conditional Knockout Mice by Sequential Insertion of Two loxP Sites In Cis Using CRISPR/Cas9 and Single-Stranded DNA Oligonucleotides
Ye Liu, Yubin Du, Wen Xie, Fan Zhang, Douglas Forrest, and Chengyu Liu
12. Improvement of Mouse Cloning from Any Type of Cell by Nuclear Injection
Sayaka Wakayama, Satoshi Kishigami, and Teruhiko Wakayama
13. The CARD Method for Simple Vitrification of Mouse Oocytes: Advantages and Applications
Naomi Nakagata, Jorge Sztein, and Toru Takeo
14. The CARD Method for Mouse Sperm Cryopreservation and In Vitro Fertilization Using Frozen-Thawed Sperm
Toru Takeo, Jorge Sztein, and Naomi Nakagata
15. Isolation and Analysis of a Genome-Edited Single-Hepatocyte from a Cas9 Transgenic Mouse Line
Takayuki Sakurai, Akiko Kamiyoshi, Masato Ohtsuka, Channabasavaiah B. Gurumurthy, Masahiro Sato, and Takayuki Shindo
16. Microinjection and Oviduct Transfer Procedures for Rat Model Generation with CRISPR-Cas9 Technology
Tuija M. Alcantar and Xin Y. Rairdan
17. Molecular Aspects of Zinc Finger Nucleases (ZFNs)-Mediated Gene Editing in Rat Embryos
Xiaoxia Cui
18. Organ Generation from Knocked-Out Rat Blastocysts Complemented with Pluripotent Stem Cells
Masumi Hirabayashi and Shinichi Hochi
19. Generation of Rabbit Models by Gene Editing Nucleases
Dongshan Yang, Jie Xu, and Y. Eugene Chen
20. Production of Genetically Engineered Porcine Embryos by Handmade Cloning
Rong Li, Jinxin Miao, and Zhongde Wang
21. Electrofusion of 2-Cell Embryos for Porcine Tetraploid Embryo Production
Qingran Kong and Zhonghua Liu
22. Gene Knockouts in Goats Using CRISPR/Cas9 System and Somatic Cell Nuclear Transfer
Zhiqiang Fan, Min Yang, Misha Regouski, and Irina A. Polejaeva
23. Generating Goat Mammary Gland Bioreactors for Producing Recombinant Proteins by Gene Targeting
Liyou An, Lan Yang, Yuejin Huang, Yong Cheng, and Fuliang Du
24. Production of Transgenic Chickens Using Cultured Primordial Germ Cells and Gonocytes
Ellen J. Collarini, Philip A. Leighton, and Marie-Cecile van de Lavoir
25. Using Microinjection to Generate Genetically Modified Caenorhabditis elegans by CRISPR/Cas9 Editing
Jyoti Iyer, Nicole DeVaul, Tyler Hansen, and Ben Nebenfuehr
26. Microinjection in Zebrafish for Genome Editing and Functional Studies
Wuhong Pei and Shawn M. Burgess
27. Microinjection of Marine Fish Eggs
Rie Goto, Taiju Saito, Takahiro Matsubara, and Etsuro Yamaha
28. Generating Gene Knockout Oryzias Latipes and Rice Field Eel Using TALENs Method
Daji Luo, Ke Feng, Zuoyan Zhu, and Wei Hu
29. Functional Studies of Transcriptional Cofactors via Microinjection-Mediated Gene Editing in Xenopus
Yuki Shibata, Lingyu Bao, Liezhen Fu, Bingyin Shi, and Yun-Bo Shi
30. Microinjection of Live Mammalian Cells: A Delivery Method that Provides Added Versatility to the Study of Cellular Function
Robert F. Gahl
This detailed book explores how microinjection will be used in the foreseeable future, not only for generating animal models for biomedical research but also for changing economically or ecologically important species that can broadly impact our society in general. The opening half of the book focuses on methods for generating mouse models, as they are still the most popular in genome engineering research, while the second half examines gene-editing in a variety of other species, opened up by the developments in ZFN, TALEN, and CRISPR techniques. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Microinjection: Methods and Protocols serves as an ideal guide for researchers looking to take advantage of the breakthrough technologies in gene-editing and embryo micromanipulations.