Synthetic far-red light-mediated CRISPR-dCas9 device for inducing functional neuronal differentiation

Area of Science:

• Synthetic biology
• Optogenetics
• CRISPR gene editing

Background:

• Precise spatiotemporal control of gene expression is crucial for understanding transcriptional dynamics.
• Optogenetic tools with deep tissue penetration and minimal phototoxicity are needed for cellular control and translational research.

Purpose of the Study:

• To engineer a far-red light (FRL)-activated CRISPR-dCas9 effector (FACE) system.
• To enable precise spatiotemporal control of endogenous gene expression using FRL stimulation.
• To demonstrate the system's utility in epigenetic modulation and cell differentiation.

Main Methods:

• Utilized synthetic biology and optogenetic design principles.
• Engineered a CRISPR-dCas9 effector activated by far-red light (FACE).
• Applied the FACE system for transcriptional activation and epigenetic modulation.

Main Results:

• Developed a versatile FACE system for FRL-inducible gene transcription.
• Demonstrated robust spatiotemporal control of endogenous gene expression.
• Successfully mediated epigenetic modulation to promote induced pluripotent stem cell differentiation into neurons via NEUROG2 upregulation.

Conclusions:

• The FACE system offers a robust and convenient method for precise spatiotemporal control of gene expression.
• FACE facilitates targeted epigenetic modulation, showing potential in cell differentiation for regenerative medicine.
• This technology holds promise for genetic/epigenetic reprogramming in biological research and biomedical applications.