Use of a Novel, Stable Gene Silencing Technology to Determine the Contribution of the Receptor Tyrosine Kinase to the Breast Cancer Phenotype
Abstract
An enormous amount of evidence has implicated members of the receptor protein tyrosine kinase (RPTK) family in many types of cancer. We have developed an extraordinarily powerful technique that uses small, DNA-encoded RNAs to suppress the expression of a desired gene in mammalian cells. For this project, short hairpin RNAs targeting each of the 58 human receptor protein tyrosine kinases will be created, introduced into MCF-7 cells and tested for effects on tumorigenicity in vivo and in vitro. At the end of the initial twelve months of the project there has been good progress. The goal of year one was to create a set of encoded hairpins targeted against the genomic complement of RPTKs. This goal has been met. By adopting high-throughput approaches to the construction of hairpins we have created a set of silencing agents not only for the genomic complement of receptor tyrosine kinases, but also all tyrosine kinases and a large number of genes functionally related to the RPTKs. Other developments include the use of shRNAs in the production of transgenic animals and microarray-based shRNA delivery for phenotypic screening. Both of these are potentially useful in future studies of the tyrosine kinase targeting constructs.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2004
- Accession Number
- ADA435144
Entities
People
- Douglas S. Conklin
Organizations
- State University of New York at Albany