GUILD ASSOCIATES INC — Department of Health and Human Services SBIR Phase I: 100

Description

DESCRIPTION provided by applicant The RNA interference RNAi mediated gene silencing has become an important technology for manipulating cellular phenotypes mapping genetic pathways and discovering therapeutic targets and has therapeutic potential A key challenge to realizing the broad potential of RNAi technology is the need for safe and effective methods for delivery Small interfering RNA siRNA into targeted cell Although a variety of materials and approaches have been explored for siRNA delivery the performance of current materials and approaches remain unsatisfactory because they generally do not work well with primary cells or non adherent cell types and often result in some degree of cytotoxicity and alterations in cell biology thereby severely limiting the cell types amenable to discovery research and not suitable for human therapeutics In this project we propose to investigate a novel molecularly designed multi functional DNA Origami nanostructures DONs for effective delivery of siRNA inside targeted cells Our hypothesis is that the biocompatible molecularly defined monodispersed DONs incorporated with siRNAs and cell penetrating peptides CPP would enables effective siRNA uptake by cells and efficient release of siRNA inside cells resulting in high gene knockdown even when using low siRNA concentrations Our ultimate goal is to develop a safe and effective DONs siRNA therapeutic for diseases the short term goal of this Phase I project is to develop DONs based multi functional siRNA delivery and transfection reagent with optimal structure to aid siRNA silencing research The Phase I proof of concept demonstration project will focus on developing chemistry and protocol for constructing DONs siRNA and in vitro assessing the effectiveness of DONs siRNA for cell transfection and gene knockdown We plan to develop one pot approach to create multi functional DONs incorporated with firefly luciferase Luc siRNA gene and green fluorescent protein GFP siRNA gene and evaluate their silencing efficiency to Luc expressed HeLa cells and GFP expressed Human prostate carcinoma PC cells In Phase II we will thoroughly optimize the preparation of multi functional DON siRNA incorporated with cell target ligands investigate the in vivo delivery of DON siRNA and the effectiveness of gene knockdown the pharmacokinetic profile and organ biodistribution of DON siRNA in nude mice bearing tumors PUBLIC HEALTH RELEVANCE In this project we plan to use biological and biocompatible natural material the DNA as building blocks to create defined DNA nanostructures as delivery carriers for short interference RNA siRNA and test the effectiveness for gene knockdown in cells The successful completion of this project is expected to develop a new type of non immunogenetic and non cytotoxic cell transfection reagent which will have broad applications in biological biomedical and pharmaceutical research as well as potential for therapeutic drug development