Gene Knockout Protocols

1. Front Matter

   Pages i-xi

   PDF

2. Overview

   • Paul J. Hertzog, Ismail Kola

   Pages 1-10

3. Isolation and Maintenance of Primate Embryonic Stem Cells

   • Vivienne S. Marshall, Michelle A. Waknitz, James A. Thomson

   Pages 11-18

4. Gene Targeting in ES Cells

   • Thomas M. DeChiara

   Pages 19-45

5. Manipulating Mouse Embryonic Stem Cells

   • Lino Tessarollo

   Pages 47-63

6. Gene Targeting in a Centralized Facility

   • Louise D. Barnett, Frank Köntgen

   Pages 65-82

7. The LoxP/CRE System and Genome Modification

   • Trevor J. Wilson, Ismail Kola

   Pages 83-94

8. Creation and Use of a Cre Recombinase Transgenic Database

   • Andras Nagy, Lynn Mar

   Pages 95-106

9. Choice of Mouse Strains for Gene Targeting

   • Mireille H. Lahoud, Trevor J. Wilson, Paul J. Hertzog

   Pages 107-119

10. Isolation, Microinjection, and Transfer of Mouse Blastocysts

    • Angelika Bonin, Susan W. Reid, Lino Tessarollo

    Pages 121-134

11. Aggregation Chimeras

    • Mika Tanaka, Anna-Katerina Hadjantonakis, Andras Nagy

    Pages 135-154

12. How to Study Pathologic Phenotypes of Knockout Mice

    • Roderick T. Bronson

    Pages 155-180

13. Analysis of Hematopoietic Phenotypes in Knockout Mouse Models

    • Ivan Bertoncello, Brenda Williams

    Pages 181-203

14. Isolation of Embryonic Fibroblasts and Their Use in the In Vitro Characterization of Gene Function

    • Paul J. Hertzog

    Pages 205-215

15. Influence of Genetic Background on Knockout Mouse Phenotypes

    • L. Philip Sanford, Suhas Kallapur, Ilona Ormsby, Thomas Doetschman

    Pages 217-225

16. Lineage Allocation During Early Embryogenesis

    • Gabriel A. Quinlan, Bruce P. Davidson, Patrick P. L. Tam

    Pages 227-250

17. Generation of Double-Knockout Embryonic Stem Cells

    • Hein te Riele, Conny Brouwers, Marleen Dekker

    Pages 251-262

18. In Vitro Differentiation of Embryonic Stem Cells and Analysis of Cellular Phenotypes

    • Anna M. Wobus, Kaomei Guan, Uta Pich

    Pages 263-286

19. Embryonic Stem Cells in the Study of Hematopoiesis

    • Francesca Lazner, Ismail Kola, Elizabeth Stadler, Andrew G. Elefanty

    Pages 287-300

20. Interferon-Inducible ES Cell Expression Systems

    • Linda M. Whyatt, Peter D. Rathjen

    Pages 301-318

As the major task of sequencing the human genome is near completion and full complement of human genes are catalogued, attention will be focused on the ultimate goal: to understand the normal biological functions of these genes, and how alterations lead to disease states. In this task there is a severe limitation in working with human material, but the mouse has been adopted as the favored animal model because of the available genetic resources and the highly conserved gene conservation linkage organization. In just of ten years since the first gene-targeting experiments were p- formed in embryonic stem (ES) cells and mutations transmitted through the mouse germline, more than a thousand mouse strains have been created. These achievements have been made possible by pioneering work that showed that ES cells derived from preimplantation mouse embryos could be cultured for prolonged periods without differentiation in culture, and that homologous rec- bination between targeting constructs and endogenous DNA occurred at a f- quency sufficient for recombinants to be isolated. In the next few years the mouse genome will be systematically altered, and the techniques for achi- ing manipulations are constantly being streamlined and improved.

"This is a valuable reference work, which will no doubt find a home on many lab benches." -E-STREAMS

• National Vision Research Institute of Australia, Carlton, Australia

  Martin J. Tymms

• Centre for Functional Genomics and Human Disease, Institute of Reproduction and Development, Monash University, Clayton, Australia

  Ismail Kola

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