Research

Research

Mission:

The Transgenic Core’s main mission is to keep up with the latest advancements in genome engineering technologies and to provide state-of-the-art services to assist NIH scientists in generating genetically engineered animal models. 

Description: 

In the past several years, the Core has successfully used the ZFN, TALEN, and CRISPR methods to generate gene-targeted mouse lines. The revolutionary CRISPR technology has enabled the Core to simultaneously target multiple genomic loci and achieve gene knockout in difficult mouse strains, such as immunocompromised mice. Besides developing these new technologies, the Core is continuing to provide a variety of services using the classical mouse genetic and reproductive methodologies, such as producing transgenic lines using the pronuclear microinjection method, generating knockout mice using ES cell-mediated homologous recombination and blastocyst microinjection, cryopreserving and resurrecting mouse lines. The Core has imported the TARGATT mouse line, which enables the insertion of a single copy transgene into a predefined genomic locus. In addition, the core also offers services for injecting stem cells or differentiated cells into immunocompromised mice for testing their ability to form teratomas or evaluating their differentiation capabilities.

Major Services:

  • Creating transgenic/knockout/knockin/conditional knockout animal models
  • Conducting gene-targeting experiments using the conventional homologous recombination or the new ZFN, TALEN, and CRIPSR methods
  • Deriving ES/iPS cells from mouse and other animal species
  • Injecting stem cells or differentiated cells into immunocompromised mice for transplantation study or tumor formation, including teratoma assays.
  • Performing IVF, mouse line re-derivation, embryo microinjection, and other assisted reproductive procedures

The Transgenic Core is located in Building 10, Room 6C103C.

Published Books and Video Publications

Microinjection Methods and Protocols book coverMicroinjection - Methods and Protocols

Series: Methods in Molecular Biology

  • Includes cutting-edge techniques for the study of microinjection and its purposes
  • Focuses on mouse models for half of the book, with the other half centered on a wide variety of other models
  • Contains key implementation advice from the experts

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.

lab worker in white coat

Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction

Zhou, D, Tan L, Li J, Liu, T, Hu Y, Li Y, Kawamoto S, Liu C, Guo S, Wang, A.

J. Vis. Exp. 2018;(141), e58467, doi:10.3791/58467

Publications