Repairing RNA Transcripts that Mediate Breast Cancer Susceptibility
Abstract
A number of genetic mutations that predispose individuals to cancer are known. For example, over 25 insertion mutations have been identified in BRCA1 and p53 transcripts that have been linked to breast cancer susceptibility. Each mutation impairs the functionality of the resultant tumor suppressor protein. Developing technology to specifically excise these mutations, thus restoring tumor suppressor activity, would be of considerable importance for the development of new molecular-based therapeutics. To this end, we have developed a novel biomolecule (a ribozyme) that can specifically excise regions from RNA transcripts. In this work, we designed a ribozyme that excises an insertion mutation that is linked to breast cancer predisposition from a short mimic of the p53 transcript in a cell-free system. We have analyzed the molecular recognition properties of this ribozyme binding substrate, which has aided our design of more effective ribozymes. Furthermore, we developed a Green Fluorescent Protein system to demonstrate that such ribozyme can excise designated insertion mutations from transcripts within bacterial cells. This ribozyme is not toxic to the cell, and appears to work with a reasonable degree of specificity. The successes in this study represent the foundation for a completely novel, yet simple molecular-based therapeutic strategy: repair disease-causing RNA transcripts by removing their insertion mutations, thereby restoring the proper function of the resultant biological products.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2005
- Accession Number
- ADA443707
Entities
People
- Stephen M. Testa
Organizations
- University of Kentucky