Molecular Biology Core

Personnel

Overview

Many of the areas of substantive research needed to understand IBD are most powerfully explored using molecular biological tools. A central goal of the Molecular Biology Core is to support studies defining molecular alterations associated with IBD, especially those related to epithelial function and immune regulation. These investigations capitalize on animal models offered by the Genetic Animal Model Core and IBD patients, and pathological specimens provided by the Clinical/Tissue Core.

In view of the existing use of molecular biological techniques in most of the laboratories of individual Center investigators, we have designed this Core to serve as a resource for guiding application of more advanced techniques. However, for those laboratories with limited expertise in molecular biology, the Core has developed a major educational effort which has proven highly popular and effective. Shared facilities and resources allow the Core to provide selected services and reagents on a more economical basis than would otherwise be possible.

Services

Molecular Biology Core provides many services to the members of CSIBD which can be divided into two group. The reagent services include

• maintenance and distribution of ES cell lines
• preparation, storage and distribution of cDNA probes, plasmid, BAC and other specialized vectors for gene expression, protein expression, and generation of transgenic and gene knock-out / knock-in mice
• ES cell gene manipulation and transfection
• storage and distribution of a wide variety of biological reagents, including eukaryotic and bacterial cell lines, ES cells, viral vectors, phage stocks, RNAs purified from tissues and cell cultures, genomic DNA for chromosomal localization, gene-specific oligonucleotides and reagents for real-time PCR and RNA interference studies, and rarely-used nucleic acid-modifying enzymes
• maintenance and distribution of selected tissue-specific Cre-expressing mice for activating gene targeting constructs in vivo
• production and distribution of thermostable DNA polymerases for standard and long PCR, as well as reverse transcription-PCR of RNA sequences
• generation of novel lines of transgenic mice

Educational services, described in more detail below, include a variety of activities to facilitate technology transfer to Center laboratories and personnel, including

• design of constructs for protein expression and mutagenesis, as well as a large variety of gene expression, transgenic and gene targeting studies
• design of primers for real time PCR and RNA interference analysis
• methods of tissue isolation, storage and preparation and labeling of RNA for microarray analysis
• use of the wide variety of molecular biological databases and analysis software (including statistical software for interpreting DNA microarray studies)

Education

Each summer Molecular Biology Core conducts a one-month course in Current Techniques in Molecular Biology for Center investigators and their staff and trainees. This program emphasizes the uses to which modern research tools can be put to explore broader biological questions. The specific topics covered evolve each year in response to developing technologies and the interests of the students. This course and associated printed and web-based materials are designed to present a broad overview of the utility of various technologies, as well as key issues in their effective use. They also include step-by-step guides to the effective use of related services provided by the CSIBD MB Core and relevant institutional cores.

In response to a high demand for supervised technology transfer the Core has organized sessions that allow selected investigators from two or three laboratories to participate simultaneously. These programs have been particularly useful for more common laboratory techniques, such as in situ hybridization, DNA-protein binding assays, development and assay of luciferase, fluorescent protein, growth hormone or beta-galactosidase reporter constructs, establishment of stably transfected cells, and protein expression and purification methodologies. The Core runs group tutorial sessions on the design of synthetic oligonucleotides for PCR, mutagenesis, hybridization and expression screening, as well as tutorials on the use of computer software and on-line services for DNA and protein sequence retrieval, sequence analysis, analysis of DNA microarray data, transgenic and knockout mouse models, tissue-specific Cre-expressing mice and image analysis. In addition, in collaboration with the Department of Molecular Biology at MGH, the Core has sponsored an intensive, two-week, full-time course designed for new students and postdoctoral fellows in commonly-used molecular biological techniques. It is offered in conjunction with Current Techniques in Molecular Genetics course. Because of constraints on space and faculty time, enrollment in the laboratory course is limited to eight students per year, mostly postdoctoral fellows in Center laboratories.

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