Work

Multiplexing light-inducible recombinases to control cell fate, Boolean logic, and cell patterningin mammalian cells

Year: 2025

Type: article

Abstract: Light-inducible regulatory proteins are powerful tools to interrogate fundamental mechanisms driving cellular behavior. In particular, 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. The design space for multichromatic circuits is limited by the small number of orthogonally addressable optogenetic switches and the types of effectors that can be actuated by them. We developed a library of red light-inducible recombinases and directed patterned myogenesis in a mesenchymal fibroblast-like cell line. To address the limited number of light-inducible domains (LIDs) responding to unique excitation spectra, we multiplexed light-inducible recombinases with our "Boolean logic and arithmetic through DNA excision" (BLADE) platform. Multiplexed optogenetic tools will be transformative for understanding the role of multiple interacting genes and their spatial context in endogenous signaling networks.

Source: Science Advances

DOI: 10.1126/sciadv.adt1971

Authors: Cristina Tous, Ian S. Kinstlinger, Maya E. L. Rice, Jenny Deng, Wilson W. Wong

Institution: Boston University

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $4,500

Funders: National Heart, Lung, and Blood Institute, National Institute of General Medical Sciences, National Institute of Biomedical Imaging and Bioengineering, National Center for Advancing Translational Sciences