Novel Targeting Approach for Breast Cancer Gene Therapy
Abstract
Poly(amidoamine) (PAMAM) dendrimers of 3.5 generation were first activated with N-hydroxysuccinimide (NHS) to obtain a stable PAMAM-NHS. This NHS activated PAMAM was then coupled with SV119 using 1-Ethyl-3-(3- dimethylaminopropyl)carbodiimide (EDC) coupling. Structural analysis of the conjugate was performed using Bruker 300MHZ 1H NMR spectrometer. PAMAM-NHS-SV119 was fluorescently labeled using FITC (Fluorescein isothiocyanate) and 1.2 molecules of FITC were found conjugated to one molecule of PAMAM. Dendriplexes were prepared with sigma receptor ligand-conjugated PAMAM dendrimers and p53-GFP plasmid. Dendriplex stability was assessed by agarose gel electrophoresis and preliminary experiments were performed to determine the transfection efficiency of these dendriplexes in MCF-7 cells and NCI/RES-ADR cells. To construct a therapeutic plasmid (p53) containing the heparanase promoter, PCR amplification of the heparanase promoter sequences was performed and the experiments to insert p53-heparanase fusion gene into the multiple cloning site of the GFP vector are in progress.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 30, 2009
- Accession Number
- ADA518928
Entities
People
- Srinath Palakurthi
Organizations
- Texas A&M University