Role of APAf-1Caspase-9 Pathway in Breast Tumor Development
Abstract
The Aim of the project was to assess the function of Apaf-l/Caspase-9 in tumorigenesis in the mouse. We panned to use mutant mice expressing a conditional knockout (KO) Apaf-l gene in the breast as the standard Apaf-1 KO mutation is embryonic lethal. Unfortunately, the generation of these mutant mice proved to be technically difficult and the production of the mutant mice has been delayed. Due to these problems, we requested a 1 year no-cost extension of the grant which was granted. As an alternative approach, we have obtained FOG mutant mice from Jackson laboratory which exhibit a hypomorphic Apaf- 1 allele. These mutant mice are viable and lack detectable expression of Apaf- 1 protein and the Apaf- 1 transcripts are 10-20 fold less abundant than those in wild-type mice. We have developed a system to express stably small (21-29 bp) interfering RNAs (siRNAs) targeted against Apaf- 1 transcripts. Eight hairpin oligonucleotides were designed to target four different regions of the Apaf- 1 transcript. We constructed and tested multiples constructs and identified a RNAi construct that effectively inhibit the expression of Apaf-1 protein in breast cancer cells.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2002
- Accession Number
- ADA412147
Entities
People
- Gabriel Nunez
Organizations
- University of Michigan