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A flexible toolkit for controlling gene expression in the mouse

Objective

The proposed project is dedicated to the development of a novel approach, called FLPFLEX, to manipulate gene expression in the mouse, for dissecting complex genetic pathways and for providing more accurate models of human disease. By engaging the cell's ow n transcriptional regulatory circuits to capture novel expression patterns, FLPFLEX cassette exhange allows multiple rapid manipulations in a single genomic locus, providing powerful and adaptable tools for designing increasingly complex genetic models of human physiology and pathology. In Module 1 will include development of readout vector technologies and establishment of the FLPFLEX cell library. We will develop flexible genomic insertion cassettes carrying modifications to allow recombinase-mediated cas sette exchange of effector genes of interest. The FLPFLEX vector carrying a multifunctional tag will be randomly incorporated into the genome of embryonic stem (ES) cells. Since a majority of all mouse genes are expressed in ES cells, multiple ES clones ca rrying an expressed FLPFLEX tag can be readily selected. We will sequence and identify 10,000 FLPFLEX cassette integration sites using mouse genomic information. Predicted expression patterns of the captured gene locus will be verified by manual and automa ted in situ hybridization of adult tissues. In Module 2, we will select 50 FLPFLEX clones for further characterization and focused studies in vivo. We will introduce different effector genes via cassette exchange in selected FLPFLEX cell clones. Germ line transmission of these exchange cassettes in mice will yield novel patterns for conditional effector genes such as Cre recombinase, or allelic exchange of individual mutated cDNAs of disease genes. The project is strategically designed to provide an efficie nt mechanism for generating, characterising and disseminating these models. Information and reagents generated will be disseminated on a publicly accessible database. '

Call for proposal

FP6-2003-LIFESCIHEALTH-I
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Coordinator

EUROPEAN MOLECULAR BIOLOGY LABORATORY
Address
Meyerhofstrasse 1
Heidelberg
Germany

Participants (4)

FONDAZIONE TELETHON
Italy
Address
Via Guglielmo Saliceto, 5
Rome
GSF - NATIONAL RESEARCH CENTER FOR ENVIRONMENT AND HEALTH, GMBH
Germany
Address
Ingolstaedter Landstrasse 1
Neuherberg
ISTITUTO DI RICERCHE DI BIOLOGIA MOLECOLARE P. ANGELETTI
Italy
Address
Via Pontina Km. 30,600
Pomezia (Rm)
HELMHOLTZ-ZENTRUM FÜR INFEKTIONSFORSCHUNG GMBH
Germany
Address
Inhoffenstrasse 7
Braunschweig