Phenotypic Knockout of Cyclin E in Breast Cancer Cells by Novel Intrabodies
Abstract
In this project, we are using a novel intrabody approach which we have recently developed to phenotypically knock-out cyclin E in order to determine the critical role of cyclin E in establishing and maintaining the transformed phenotype of breast cancer cells, and to demonstrate whether cyclin E can be used as a therapeutic target. In last 10 months, the variable region cDNA genes of two anti-cyclin E antibodies against the cyclin box domain have been cloned, and sequenced. The cDNA genes were assembled into single-chain antibodies (sFv) consisting of immunoglobulin heavy- and light-china variable domains joined by a flexible peptide linker. The sFv fragments were then cloned into various expression vectors. We are in the process to characterize the binding activities of anti-cyclin E sFvs by using a novel recombinant phage antibody system. We will generate breast cancer cell lines expressing the anti-cyclin E intrabodies to examine the ability of the anti-cyclin E intrabodies to bind cyclin B in vivo and their effects on the morphology, growth properties and cell cycle progression of untransformed human fibroblasts and breast cancer cells.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 1998
- Accession Number
- ADA353778
Entities
People
- Si-yi Chen
Organizations
- Wake Forest School of Medicine