# Advanced recombinase-based gene expression technology in mammalian cells

> **NIH NIH R01** · BOSTON UNIVERSITY (CHARLES RIVER CAMPUS) · 2022 · $330,000

## Abstract

Project Summary/Abstract
Gene expression, together with genome engineering, technology are the cornerstones of the biotechnology
revolution. While much advances have been made, most gene expression control systems were designed to
control a single gene. However, many biological processes, such as developmental and cancer progression,
are driven by the simultaneous changes of expression for multiple genes in a sequentially and spatially
controlled fashion. As our genome editing capabilities rapidly progress, the development of gene expression
control technology has fallen behind. Advancement in gene expression technology that enables multiplexed
spatiotemporal control is therefore urgently needed to directly interrogate complex biological processes and to
engineer novel phenotype for biotechnological applications.
Site-specific DNA recombinase (SSR) (e.g., Cre and Flp) has become one of the most powerful gene
regulation tools in mammalian cells. We and others have shown that recombinases are uniquely capable of
creating exceptionally complex logic circuits with high robustness. As such, recombinase represents an ideal
foundation for engineering advanced gene expression control systems. Most of the recombinase-based
expression technologies were designed using one enzyme, Cre, which affords limited functionality concerning
the number of genes that can be regulated independently in a spatiotemporal manner. To develop the next
generation of recombinase-based gene expression technology, robust orthogonal inducible recombinases are
necessary. Leveraging our vast experience in engineering inducible recombinases and recombinase-based
circuit, we will develop a suite of advanced recombinase-based tools that show simultaneous, sequential,
and/or spatially controlled tuning of the expression of multiple chosen genes. In particular, we will
Aim 1: Develop orthogonal small molecule inducible recombinases for simultaneous control of
multiple gene expressions independently in the same cell.
Aim 2: Develop multichormatic light inducible gene switches for spatial control of gene expression
Aim 3: Develop cascade circuits for sequential control of gene expression
We will validate our system in human and mouse cells to ensure broad applicability. We will develop metric
and datasheet, a database for DNA repository to facilitate adoption and sharing. My group is uniquely capable
of accomplishing this proposed work because of our published expertise in 1) DNA recombinases and 2)
genetic circuit designs. Success from this proposed work will dramatically increase our capability to control
gene expression in mammalian cells with enhanced spatiotemporal precision.

## Key facts

- **NIH application ID:** 10350656
- **Project number:** 5R01GM129011-04
- **Recipient organization:** BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
- **Principal Investigator:** Wilson Wong
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $330,000
- **Award type:** 5
- **Project period:** 2019-06-01 → 2024-02-29

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10350656

## Citation

> US National Institutes of Health, RePORTER application 10350656, Advanced recombinase-based gene expression technology in mammalian cells (5R01GM129011-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10350656. Licensed CC0.

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