Functional Genomics for Epithelial-Mesenchymal Transition in Breast Cancer
Abstract
To assess whether modulation of the PMC42-LA cell line by a library of shRNAmir constructs can cause tumorigenicity, the impact of a smaller boutique library is first being assessed, as proposed. This boutique library of 4,462 shRNA constructs targeting 1,860 markers and mediators of EMT, metastasis, migration, and BCSC has been transduced into the PMC42-LA cell line as a total pool and as ten smaller sub-pools. These were expanded in preparation for in vivo testing. The smaller sub-pools have greater representation of individual shRNAmir constructs. For any sub-pool containing shRNAmir constructs that enables tumorigenicity, this greater representation translates to a markedly larger inoculum of tumorigenic cells, which may be important in the establishment of a tumor. The presence of a small but significant proportion of colonies exhibiting markedly mesenchymal features has been observed in the transduced pools, validating the use of this boutique library pool to directly assess whether PMC42-LA tumors can be grown using pools containing a proportion of more mesenchymal cells as an inoculum prior to assessing the whole genome shRNAmir library. Various methodologies for the in vitro selection of transduced cells with mesenchymal features from these pools, supplementary to the initial aims, are under development.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2011
- Accession Number
- ADA554588
Entities
People
- Erik Thompson
Organizations
- University of Melbourne