Project Coordinator - Lisa Hines                                                           Current List of Genes
Research Assistant - Donna Martins
Research Assistant - Jacob Raphael

BREAST CANCER 1000 PROJECT (BC1000) 

The objective of the Breast Cancer 1000 Project (BC1000) is to understand key aspects of breast development and breast carcinogenesis.  The principal research emphasis is on identifying and understanding the biological function of proteins that play important roles in controlling these processes, which can be achieved through the development and availability of high-throughput protein expression methodologies.  Our goal is to complete and distribute a ready-to-use repository of clones for 1000 full-length cDNAs for genes associated with breast cancer development and breast carcinogenesis.  The BC1000 gene list is comprised of genes suggested by our colleagues and genes identified using MedGene, a text-mining software developed at the Harvard Institute of Proteomics (HIP). 

      This project exploits recombinational cloning technology, which allows DNA fragments flanked by site-specific recombination sites to be moved easily from one vector to another.  We are capturing the genes into the Creator™ recombination system using the BD In-Fusion™ Cloning Kit (BD Biosciences) to develop a useful library of full-length coding sequences of genes involved in breast development and breast carcinogenesis.  This novel gene cloning strategy allows DNA fragments surrounded by site specific recombination sites to be moved easily from one vector to another.  HIP has developed protocols to adapt this gene cloning strategy to an automated and high-throughput format.  Once these “master clones” are assembled and verified, this set of genes can be readily transferred into any protein expression vector to enable the widest possible range of functional experimentation such as in vitro translation assays, kinase substrate screens, or in vivo phenotypic rescue experiments. 

      To complement this collection, we will develop the technology needed to facilitate functional experimentation with the BC1000.  This will include the modification of useful expression vectors and the development of automated methods for high-throughput transfer of these clones to these vectors for use in molecular biology, functional, or genetic screens.  In addition, we will develop and maintain a database for the completed clones and potentially the results obtained from the experiments done with the BC1000.  This database can provide a central location to query for available clone information and to stimulate new research ideas from the existing experimental results.