Microinjection: Methods and Protocols

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 TARGATT^TM 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

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

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