# Multiplexed Optogenetic Control of Mammalian Genome and Transcriptome using Recombinases and Cas13

> **NIH NIH F31** · BOSTON UNIVERSITY (CHARLES RIVER CAMPUS) · 2024 · $35,139

## Abstract

Abstract:
Light-inducible regulatory proteins are powerful tools to interrogate the fundamental mechanisms driving
cellular behavior. To this end, genetically encoded photosensory domains fused to split proteins can tightly
modulate protein activity and gene expression. While light-inducible split protein systems have performed well
individually, few multichromatic and orthogonal gene regulation systems exist in mammalian cells. Existing
multi-input circuits are hampered by their type of regulation and the scale of possible outputs given the number
of wavelengths. I will address this limitation by creating a library of red and blue light-inducible split
recombinases and the first suite of split Cas13 ribonucleases. Site specific recombinases provide permanent
and transgene outputs, while Cas13 ribonucleases will provide a complementary approach to modulate
reversible analog transcriptomic outputs. The multiplexed optogenetic tools developed in this proposal will be
transformative for understanding the role of multiple interacting genes in endogenous signaling networks. I will
leverage the ability of C2C12 myoblasts to differentiate into osteoblasts or myotubes and encode an
illumination-dependent cell fate switch. While C2C12 cells have been a fundamental model in studying how
differentiation is impacted by mechanical cues and growth factors, this work will explore spatiotemporal control
of optogenetic regulatory proteins in order to direct cell fate.

## Key facts

- **NIH application ID:** 10884216
- **Project number:** 5F31GM149163-02
- **Recipient organization:** BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
- **Principal Investigator:** Cristina Tous
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $35,139
- **Award type:** 5
- **Project period:** 2023-07-01 → 2025-04-22

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10884216

## Citation

> US National Institutes of Health, RePORTER application 10884216, Multiplexed Optogenetic Control of Mammalian Genome and Transcriptome using Recombinases and Cas13 (5F31GM149163-02). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10884216. Licensed CC0.

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