# Next generation transposon vectors for genome engineering

> **NIH NIH R01** · VANDERBILT UNIVERSITY MEDICAL CENTER · 2022 · $525,492

## Abstract

Non-viral gene delivery systems are limited by their activity and targeted integration capability. Efficient and
targeted integration of DNA into mammalian and human genomes remains a major challenge and its success
would have wide impact for biotechnology and therapeutic applications. The piggyBac (PB) transposon system
is the most active integrating non-viral gene delivery system and is a cut-and-paste DNA transposon that
has been used for genome engineering of mammalian and human cells for more than 15 years. We
have re-engineered the PB-transpososome (transposase with transposon DNA) based on the first-
ever three-dimensional structure of the PB transpososome that we recently published with our collaborator
Dr. Fred Dyda (Chen et al., Nature Communications, 2020). Our next-generation PB transpososome
(ngPB) demonstrates greater activity and potential for targeted integration than was previously achievable. In
specific aim 1, we will engineer and test ngPB for genome engineering of human cells. We will evaluate the
integration site profile and copy number of transposon integrations per human cell. We will modify primary
human T cells ex vivio and test their ability for cell therapy, and we will enable transposase protein transfection.
In specific aim 2, we will engineer and test ngPB for gene delivery in vivo. We will evaluate gene delivery of
reporter and therapeutic transgenes to mouse liver, test for efficiency in development of transgenic mice, and
evaluate hybrid adeno-associated viral (AAV)-ngPB mediated gene delivery to difficult to reach organs. In
specific aim 3, we will engineer and test ngPB for targeted integration in human cells. We will also map the
protein-protein interaction domain of PB known to affect its target site selection in human cells and test PB
protein modifications to allow greater flexibility in manipulating PB genomic target site selection. The
proposed studies will be transformative for genome engineering and have broad impact for biotechnology
and therapeutic applications.

## Key facts

- **NIH application ID:** 10501335
- **Project number:** 1R01EB033676-01
- **Recipient organization:** VANDERBILT UNIVERSITY MEDICAL CENTER
- **Principal Investigator:** MATTHEW H WILSON
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $525,492
- **Award type:** 1
- **Project period:** 2022-09-01 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10501335

## Citation

> US National Institutes of Health, RePORTER application 10501335, Next generation transposon vectors for genome engineering (1R01EB033676-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10501335. Licensed CC0.

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