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2. Use of genetically modified mice in biomedical research
3. Core B Function One of the greatest obstacles for investigators who wish to use mouse genetics to study their gene of interest is in the generation of DNA-targeting constructs.
The Molecular Engineering Core will function to generate targeting cassettes for the production of both transgenic and knockout animals
*(Core B does not perform ES electroporation, blastocyst injections or DNA microinjections- see Dr. Bob Kesterson at the UAB transgenic mouse facility for these services)
To start a project, go to website and fill out job request form
4. Core B Services Advise participating investigators on the generation and design of DNA constructs that are optimal for their studies
Construct the necessary targeting cassettes and prep DNA for injections or electroporation
3. Assist in the development of genotyping protocols for the maintenance of new mouse lines.
5. Transgenic Constructs
6. Constitutive gene-inactivation constructs Gene-trap alleles
Constitutive inactivation
Tagged with B-gal for use in determining expression patterns
ES cells libraries already generated
database search identifies possible targeted genes
Bay genomics/MMRRC
allele may be null or hypomorphic
7. Constitutive gene-inactivation constructs Constitutive gene knockout
Production of gene-replacement cassettes
Identify BACs encoding gene
Retrieve genomic fragments and replace exons with the ES cell selectable marker neomycin
Sequence construct to confirm correct replacement
Prep DNA for electroporation into ES cells
8. Conditional gene-inactivation constructs Conditional gene knockout using Cre-lox methodology
Two components
insertion of loxP sites into a neutral region surrounding critical exon/s
expression of Cre recombinase
Allows for gene inactivation in a restricted neuronal population and at desired time in development
Construct floxed allele
Sequence confirm and prep DNA for electroporation
12. Genotyping The core will develop PCR based genotyping protocols for DNA targeting cassettes
Both standard PCR and qPCR genotyping available
Provide assistance and information on maintenance of mouse colony
breeding, colony size and maintenance, book keeping
tail clip, ear punch, mating, weaning, chow
13. Core B- Molecular Engineering Personnel
Open Position- Technical Director
Sylvia Huang- Research Assistant
Ludwig Francillion- Laboratory Assistant
Contact Info
Scott Wilson
5-5573
SHELBY 972