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

> **NIH NIH R43** · GRADIENT MEDICAL, INC. · 2021 · $251,914

## Abstract

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.

## Key facts

- **NIH application ID:** 10324621
- **Project number:** 1R43GM143999-01
- **Recipient organization:** GRADIENT MEDICAL, INC.
- **Principal Investigator:** Jordan Fong
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $251,914
- **Award type:** 1
- **Project period:** 2021-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10324621

## Citation

> US National Institutes of Health, RePORTER application 10324621, Feasibility of INSPIRE Technology to Enhance CRISPR Mediated Genome Editors In Vivo (1R43GM143999-01). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10324621. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*