Restore Wild-Type Functions to P53 Mutants Using an RNA- Based Combinatorial Approach
Abstract
The tumor suppressor p53 plays pivotal roles in maintaining the integrity of the genome and regulates cell cycle arrest and programmed cell death (apoptosis). Tumors without a functioning p53 are more likely to have a very aggressive clinical course. We proposed to use an RNA-based combinatorial approach to select for single-stranded RNA molecules that will bind to a mutant p53 protein, correct its conformation, and restore its sequence-specific DNA-binding activity. We have successfully accomplished the tasks we proposed for the first year of funding, including (1) expression and purification of human wild-type p53 protein in insect cells, (2) in vitro selection of RNA aptamers from a pool of RNA molecules that bind to p53 protein, and (3) sequence-specific DNA binding assays using the EMSA technique. Sequence and structural analysis of the selected RNAs revealed that the ability of RNA molecules to interfere with sequence-specific DNA binding of p53 may depend on their tertiary structures.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 1998
- Accession Number
- ADA353779
Entities
People
- Xiaoying Chen
Organizations
- SRI International