Genetic Resource Science

Technology Project: Genetic Tools and Resources for

Orofacial Clefting Research

Leah Rae Donahue, Ph.D.Stephen Murray, Ph.D.

Overall goal of this project

To facilitate orofacial clefting research• by generating new mouse genetic tools• by providing a repository of mouse strains

critical for the clefting research community

Mouse is a useful phenotypic and genetic model

Genetic Resource Science

Richtsmeier et. al., 2000. Devel Dyn 217 (2):138

Genetic Resource Science

Aims

1. To generate 15 new, well-characterized Cre driver lines to facilitate orofacial clefting research.

2. To provide a repository for importation, cryopreservation, GQC, and distribution of mouse models and tool strains important for orofacial clefting research.

Aim 1:

To generate Cre driver lines

Genetic Resource Science

•That the generation of inducible Cre (CreERT2) drivers where practical would best complement and expand the existing set of Cre strains and best facilitate in vitro experimentation.

•That a simultaneously expressed fluorescent reporter be incorporated into the design.

•That a goal of 15 Cre driver lines be proposed, of which a subset would be chosen at a later date to allow data sets generated as part of the FaceBase effort to contribute to the design of future specific driver lines.

•That the choice of driver promoters include a combination of highly conserved enhancer elements (from A. Visel and colleagues) and traditional gene promoters.

•That BAC transgenics (for traditional promoters) and ROSA26 knock-ins (for conserved elements) provide the best overall approach.

•That characterization of Cre function should be comprehensive between E11.5 and E15.5 using an established reporter (such as R26R-LacZ).

•That potential non-specific Cre activity should be carefully evaluated using the established Cre characterization pipeline at TJL.

Design Principles for the Cre Driver Project

Brain Lung Adrenal (High Mag.)Testis Kidney

Nes-Cre

I.H.G.F. J.

Brain Epididymus D. (High Mag.)KidneyOvary

ChAT-Cre

E.A. D.C.B.

Brain Heart MuscleLung Kidney

Thy1-Cre

L.K. O.M. N.

Unexpected Cre activity

Cre Repository website: direct access to strain information and characterization data

www.jax.org/jaxmice/research/cre

Recently released via MGI…Searchable database of Cre activity

Aim 2:

To provide a repository of mouse models and genetic tools for orofacial clefting research.

Genetic Resource Science

Resource management is part of the JAX tradition

• provided mouse strains and associated resources to the international scientific community for nearly 80 years

• historically through individual JAX investigators

• now through the JMS & the Genetic Resource Science Repository

•

• A collection of mouse resources ~ 4300 strains total ~ 1300 strains kept as breathing colonies ~ 3000 cryopreserved strains

• Information about them– MGI – JAXMice– MPD– Specialized web sites

The GRS Repository is…..

GRS Repository has three large resource programs

• The Mouse Mutant Resource for spontaneous mutants, federally funded for 50 years

• Induced Mutant Resource, repository for mice carrying transgenes, and targeted or chemically induced mutations, funded 1992

• The Special Mouse Strains Resource, panels of consomic and recombinant

inbred strains, funded 2001

GRS Specialized Repositories

• Craniofacial Resource• Eye Models • Hearing Models • Cytogenetic Models• Neural Tube Defects

Models• Cystic Fibrosis Resource• Type I Diabetes Resource • Alzheimer’s Disease

Models

• Parkinson Disease Models• Neuronal Tools Strains• SMA Resource• Friedreich’s Ataxia

Resource• Recent additions– Cre Repository– FaceBase, Models of

Orofacial Clefting

The FaceBase Repository at JAX

• Seek new models • Import mice into specific pathogen-free status• Cryopreserve imported and newly developed Cre driver

strains (no cost to FaceBase members)• Perform genetic quality control • Maintain live colonies for timely access to research models• Distribute mice through TJL's established distribution

program• Create and maintain the TJL FaceBase website to promote the

use of this resource and to facilitate access to mouse models and tools for orofacial research (linked to Hub, of course)