Permanent URL to this publication: http://dx.doi.org/10.5167/uzh-33423
Johansson, T; Broll, I; Frenz, T; Hemmers, S; Becher, B; Zeilhofer, H U; Buch, T (2010). Building a zoo of mice for genetic analyses: A comprehensive protocol for the rapid generation of BAC transgenic mice. Genesis: The Journal of Genetics and Development, 48(4):264-280.
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Transgenic mice are highly valuable tools for biological research as they allow cell type-specific expression of functionally instrumental genes. In this protocol, the generation of bacterial artificial chromosome (BAC) transgenic constructs is described. We give an overview of different transgenic inserts, such as fluorescent proteins (alone or in combination with Cre variants), diphtheria toxin receptor, lacZ, and light-activated ion channels. The most reliable and versatile approach to express these genes is by using BACs, which allow "highjacking" of the expression pattern of a gene without characterizing its transcriptional control elements. Here, we describe the necessary cloning techniques compared with conventional transgenesis. With the provided "toolbox" of already available transgene constructs, the generation of the BAC transgenes is made easy and rapid. We provide a comprehensive outline how to insert the different transgenes into a chosen BAC by either ET cloning or recombineering. We also describe in detail the methods to identify the correct insertion and the integrity of the final BAC construct, and finally, the preparation of the BAC DNA for oocyte injection is described. genesis 00:1-17, 2010. (c) 2010 Wiley-Liss, Inc.
|Item Type:||Journal Article, refereed, original work|
|Communities & Collections:||04 Faculty of Medicine > Institute of Pharmacology and Toxicology|
04 Faculty of Medicine > University Hospital Zurich > Institute of Experimental Immunology
|DDC:||570 Life sciences; biology|
610 Medicine & health
|Deposited On:||06 May 2010 13:08|
|Last Modified:||27 Nov 2013 18:36|
|Citations:||Web of Science®. Times Cited: 10|
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