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Cell and Molecular Biology


Objectives
The Cell and Molecular Biology Core:

Specific Services

Advice and Training in Cell and Molecular Biology Techniques
The core can provide advice regarding the design and interpretation of experiments using basic molecular techniques such as PCR, transfections, etc. The core can assist with the computer analysis of DNA or protein sequences. Users who have highly specialized needs that require expertise beyond the core will be referred to other appropriate laboratories where feasible.

Transgenic Animal and Vector Core Facility Subsidies
The MDRTC Cell and Molecular Biology Core can provide limited subsidies (typically 25% of cost, though the total subsidy for each laboratory may be capped) to users of the Biomedical Research Council Transgenic Animal and Vector Core Facilities. A Transgenic Animal Core Subsidy Application or a Vector Core Subsidy Application should be completed and submitted via the Web. The application will be reviewed, and you will be notified of approval/ disapproval. To be potentially eligible for the subsidy, the project must be relevant to diabetes or related endocrine disorders, and this relationship must be clearly justified on the application form. For the purpose of these applications, “related endocrine disorders” are those that impact diabetes, such as obesity.

Microarray Analysis
Gene expression profiling using Affymetrix GeneChips is available at a discounted rate through the Cancer Center Microarray Facility. To be potentially eligible for the subsidy, a Microarray application form should be filled out and submitted via the Web. The application will be reviewed, and you will be notified of approval/ disapproval.The project must be relevant to diabetes or related endocrine disorders, and this relationship must be clearly justified on the application form. For the purpose of this application, “related endocrine disorders” are those that impact diabetes, such as obesity. Microarray analysis is very expensive. The MDRTC typically will provide a 25% discount, though the total subsidy for each laboratory may be capped.

Hybridoma Core
The MDRTC will continue to provide subsidies for use of the Hybridoma Core, typically equal to 50% of charges, though the total subsidy for each laboratory may be capped. Please see the Hybridoma Core web site. To be potentially eligible for the subsidy, the project must be relevant to diabetes or related endocrine disorders. “Related endocrine disorders” are those that impact diabetes, such as obesity.

RNA interference / shRNA lentiviral plasmids
The UM Functional Genomics Core (3338 LSI; 615-5309; http://fgc.lsi.umich.edu) houses, maintains and distributes lentivrial plasmid vectors expressing shRNAs directed against most mouse and human genes, to be used in RNA interference gene silencing studies. The MDRTC Cell & Molecular Biology Core will provide a 50% subsidy toward the purchase of these shRNAs, though the total subsidy for each laboratory may be capped. An application form is not necessary. There should be an MDRTC check box on the UM Functional Genomics Core request form, which will trigger the subsidy.

High Throughput Chemical Screens (Center for Chemical Genomics, 3338 LSI, 615-9422, http://www.lsi.umich.edu/ccg)
The Center for Chemical Genomics performs high throughput robotics screens of a library of ~50,000 small molecules to identify compounds that activate or inhibit a biological process of interest. Typical goals are to either: a) identify novel chemicals to rapidly activate or inhibit the process and study the biological consequences; or b) find potential leads for the identification of useful drugs. The MDRTC can subsidize 1 or 2 users per year at 50% of the cost up to $10,000 per PI. (We will subsidize the actual small molecule screen, after assay development is complete). The project should be discussed with the CGC directly to assess feasibility and to develop the screening assay. The CCG form contains an “Other Funding Information” box; you apply for the subsidy by writing MDRTC in that box. To be potentially eligible for the subsidy, the project must be relevant to diabetes or related endocrine disorders. “Related endocrine disorders” are those that impact diabetes, such as obesity.

Equipment

The equipment described below is available to MDRTC members and is located in room 5570-E MSRB-II. Room keys cannot be provided to users, and the room must be kept locked when it is not in use. We regret the inconvenience, but this means that users may not have reliable access during non-business hours. Users must first receive training by Drs. Ron Koenig or Dennis Larkin before they will be able to access the equipment.

Bacterial Artificial Chromosome Recombineering Core Update

Bacterial artificial chromosome (BAC) libraries with all the genes from human, mouse and other species are commercially available. Modification of the large genomic inserts to reproduce disease-causing mutations or to mark cells expressing certain genes is done by recombineering. The Recombineering Core Facility was established in the Transgenic Core by a partnership with the Endowment for the Basic Science, the Diabetes Research and Training Center, the Cancer Center, the Gastrointestinal Peptide Center, the Life Sciences Institute and the Center for Organogenesis. The Recombineering Core has set up a wet lab for and is recombineering BACs for pilot projects. Several BACs have been successfully modified with lacZ and red fluorescent proteins. Plasmid vectors to introduce other genetic markers (green fluorescent protein and Cre recombinase) are under construction in the Core. DNA sequencing of modifications and BAC fingerprinting are used for quality control purposes. We will continuously refine our methodology as we work through pilot projects. After we recombineer a few more BACs we will open the BAC Recombineering Core to the research community at large. If you are interested in BAC services, please contact Thom Saunders (tsaunder@umich.edu).

Key Personnel

Ronald J. Koenig, M.D., Ph.D., Core Director, rkoenig@umich.edu; 734-763-3056