Development of a Nanotechnology Platform for Prostate Cancer Gene Therapy
Abstract
The objective of this research is to design and develop a nanocarrier that is able to evade the immune system, circulate in the blood stream, find its target prostate cancer cells, and transfer therapeutic genes into prostate cancer cells efficiently. The gene carrier is composed of: a) histone H2A peptide (H2A) to condense pDNA into nano-size particles (nanocarriers), b) a PC-3 specific targeting motif (TM) to target prostate cancer cells, c) an endosome disrupting motif (EDM) to disrupt endosome membrane and facilitate escape of the cargo into the cytosol, and d) a nuclear localization signal (NLS) to actively translocate pDNA towards the nucleus of cancer cells. The gene encoding the gene delivery system was synthesized and cloned into a pET21b vector. The vector was transformed into E.coli, expressed and purified. The purity (>96%) and expression of the gene delivery system was confirmed by SDS-PAGE and westernblot analysis. The gene carrier was complexed with plasmid DNA (pDNA) to form stable nanoparticles with sizes below 100nm. The nanoparticles were used to deliver reporter genes (pEGFP) to target PC-3 prostate cancer cells. The results demonstrated that the gene delivery system is able to target and efficiently transfect PC-3 cancer cells with minimum cross-reactivity with normal epithelial prostate cells. Furthermore, the gene delivery system by itself did not show any detectable toxicity in the range tested (110ug/ml).
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2010
- Accession Number
- ADA527848
Entities
People
- Arash Hatefi
Organizations
- Washington State University