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 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

&nbs