GUDMAP Reporter Strain Nominations

Generation of new mouse strains for analysis of the developing urogenital system

NIDDK has recently initiated a second round of funding for the Genito-Urinary Developmental Anatomy Project. As part of this multicenter effort, GUDMAP2 members will generate novel transgenic mouse strains using gene-specific targeting to mark (fluorescent reporter) key cell populations of broad interest to the community and to facilitate genetic modification within the cell-of-interest (tamoxifen-inducible CRE recombinase). Gene nominations for targeting are sought from the research community (see form below). GUDMAP2 will give preference to gene targets expressed in critical cell types of the lower UGS, but will consider cell-type specific marking throughout the UGS. Once generated and verified, mice will be made available to nominating investigators and deposited in a Mutant Mouse Regional Resource Centers (MMRRC) for broader distribution to the community.

Decisions for the first round of mice will be made mid-November and additional mice will be considered until the end of 2015. Further details can be found by contacting Manfred Baetscher MBaetscher@MCB.harvard.edu for technical details or Deborah Hoshizaki dkhosh@nih.gov for information about GUDMAP2.

GUDMAP2 - Gene Nominations Form

Fields marked with * are required.

Date Proposer Name * Insititute * E-Mail * ensure e-mail is correct as confirmation will be sent to this address Phone Number

Gene Symbol * Common Name MGI Gene ID *

Expression in UGS * Where and when? Expression elsewhere if known

Is there a strain currently available? Is it in a public repository?

Justification and value added to GUDMAP2 * How will this mouse benefit research of the UGS?

Other relevant information

Useful reference on candidate gene expression

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GUDMAP2 Strategy for Production of Reporter Strains

Strategy: The GUDMAP2 reporter strain production is based on the use of pre-existing targeted gene loci from the KOMP-CSD/EUCOMM ES cell clone library. ES cell clones will be modified by swapping the targeted allele with a multifunctional cassette comprising the fluorescent reporter, EGFP, and the tamoxifen-inducible recombinase, CreERT2, using a process known as dual-recombinase-mediated cassette exchange (dRMCE). Novel multifunctional reporter cassettes used with dRMCE are being developed at the Sanger Institute and mouse strains produced in GUDMAP2 will complement efforts by EUCOMMTOOLS. Following modification and verification of the modified allele, ES cell clones will be used for construction of chimeras and the derivation of novel reporter strains.

Steps: 1. Procuring ES Cell clones comprising KOMP-CSD/EUCOMM targeted loci as shown in (1). 2. Modify ES cell clones with the multifunctional cassette (2) by dRMCE. 3. Verification of modified loci by DNA sequence analysis. 4. Generation of chimeras, germ line testing and breeding F1 generation. 5. Distribution of F1 animals to nominating investigators, GUDMAP2 consortium, and MMRRC/Jax for colony expansion and archiving.

Further Considerations: 1. The objective of GUDMAP2 is to produce 5 reporter strains in year 1. Current plans call for production of 20 more strains between 2012-2016. 2. Nomination process for the first 5 genes expected to be completed by early 2012. 3. GUDMAP2 reporter cassette is currently fine-tuned and validated. Depending on functionality and efficacy, final cassette may slightly differ from cassette shown in (2).

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