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Feasibility of INSPIRE Technology to Enhance CRISPR Mediated Genome Editors In Vivo

INSPIRE 技术增强 CRISPR 介导的体内基因组编辑的可行性

基本信息

批准号：
10324621
负责人：
Jordan Fong
金额：
$ 25.19万
依托单位：
GRADIENT MEDICAL, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10324621
关键词：
3-Dimensional 3D Print Address Animal Model Animals Antibodies CRISPR/Cas technology Cell Nucleus Cell membrane Cells Charge Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collagen Computer software Custom DNA DNA Vaccines Data Defect Deoxyribonuclease I Dermal Development Devices Dose Electroporation Enzymes Epidermis Evaluation Event Family suidae Feasibility Studies Fibroblasts Gene Delivery Generations Genes Genetic Engineering Genome Guide RNA Hand Human In Vitro Injections Label Laboratories Malignant Neoplasms Mediating Messenger RNA Methods Modeling Muscle Muscle Contraction Pain Phase Physiologic pulse Plasmids Probability Process Proteins Protocols documentation Public Health Reagent Research Design Safety Series Serum Skin Subcutaneous Tissue System Techniques Technology Testing Time Tissue Viability Tissues Transfection Transistors Transmembrane Transport Transportation Vaccination Width base design efficacy study electric field endonuclease experimental study genome editing histological specimens improved in vivo in vivo Model millisecond minimally invasive nanoscale nanosecond novel portability prevent prototype silicon carbide simulation vaccine candidate vaccine delivery voltage

项目摘要

Project Summary / Abstract This applications long-term objective is to develop a platform technology enabling in vivo genetic engineering technologies including DNA vaccination and precision CRISPR gene editing. A challenge in utilizing CRISPR technologies in vivo is the need to simultaneously deliver the DNA endonuclease enzyme, guide RNA, and payload DNA. Integrated nanosecond pulse intradermal reversible electroporation (INSPIRE) is a novel technique for enhancing in vivo delivery of these components. INSPIRE rapidly transports large molecules into cells by using ultrashort electrical pulses to simultaneously induce temporary nanoscale defects in the cell membrane and drive the target molecules against concentration gradients via electrokinetic drift. This proposal will demonstrate the feasibility of INSPIRE for in vivo genetic engineering in a 3D in vitro skin model and a porcine dermal model via three specific aims. 1) Development of a Hand Held System Suitable for Clinical INSPIRE Treatments, 2) Optimization of INSPIRE Protocols in a Laboratory Model, and 3) Feasibility of INSPIRE Protocols in a Large Animal Model. In Aim 1 we will construct a portable pulse generation system suitable for clinical use with the high voltage components integrated directly into a hand held applicator. In Aim 2 we will optimize protocols for delivering CRISPR cas9 plasmid, cas9 mRNA, and cas9 protein into cells using a 3D in vitro skin mimic. In Aim 3 we will demonstrate the feasibility of delivering these CRISPR components in vivo via a porcine dermal model. Following this Phase I application demonstrating in vivo feasibility, Phase II will focus on design for manufacture and initiate long-term safety/efficacy studies designed to determine serum levels of antibodies produced following transfection with DNA vaccine candidates.

项目摘要/摘要