Gene Targeting in Normal Human Breast Epithelial Cells
Abstract
This exploration grant was to test if it is possible to achieve efficient homologous recombination and gene targeting in immortalized but otherwise normal human breast epithelial cells. Although gene targeting has been achieved in somatic human cells using transfected targeting vectors, the efficiency of this process is very low necessitating the isolation and analysis of many thousands of cell clones before a targeted allele can be identified. Such approaches are impractical with normal human breast cells. We tested if the adoption of a different delivery method, needle microinjection, resulted in more efficient homologous recombination such that targeted clones could be identified after screening fewer colonies. Although we obtained antibiotic resistant clones, we have been unable to identify any clones with targeted alleles. We conclude that our idea was incorrect and that this approach will not be practically feasible without further modification.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 01, 2004
- Accession Number
- ADA432980
Entities
People
- Andrew M. Thorburn
Organizations
- Wake Forest University