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Beta-cell-specific transgenic mice provide an invaluable model for dissecting the direct signaling mechanisms involved in regulating beta-cell structure and function. Furthermore, generating novel transgenic models is now easier and more cost-effective than ever, thanks to exciting novel approaches such as CRISPR.Here, we describe the commonly used approaches for generating and maintaining beta-cell-specific transgenic models and some of the considerations involved in their use. This includes the use of different beta-cell-specific promoters (e.g., pancreatic and duodenal homeobox factor 1 (Pdx1), rat insulin 2 promoter (RIP), and mouse insulin 1 promoter (MIP)) to drive site-specific recombinase technology. Important considerations during selection include level and uniformity of expression in the beta-cell population, ectopic transgene expression, and the use of inducible models.This chapter provides a guide to the procurement, generation, and maintenance of a beta-cell-specific transgene colony from preexisting Cre and loxP mouse strains, providing methods for crossbreeding and genotyping, as well as subsequent maintenance and, in the case of inducible models, transgenic induction.

Original publication

DOI

10.1007/978-1-0716-0385-7_13

Type

Journal article

Journal

Methods in molecular biology (Clifton, N.J.)

Publication Date

01/2020

Volume

2128

Pages

181 - 205

Addresses

Department of Diabetes, School of Life Course Sciences, King's College London, London, UK. lorna.smith@kcl.ac.uk.

Keywords

Animals, Mice, Transgenic, Mice, Integrases, Crosses, Genetic, Genetic Engineering, Organ Specificity, Gene Expression, Genes, Reporter, Insulin-Secreting Cells, Promoter Regions, Genetic, Gene Knockout Techniques, Genotyping Techniques