Light-Triggered CRISPR/Cas12a for Genomic Editing and Tumor Regression

Area of Science:

• Molecular Biology
• Gene Editing Technologies
• Biotechnology

Background:

• CRISPR/Cas12a systems offer precise gene editing capabilities.
• Controlling CRISPR/Cas12a activity spatially and temporally remains a challenge.
• Photo-responsive systems can enhance the spatiotemporal control of gene editing.

Purpose of the Study:

• To develop a photo-triggered CRISPR/Cas12a gene editing system (LAC12aGE).
• To demonstrate the efficacy of LAC12aGE for in vitro and in vivo gene editing.
• To evaluate the therapeutic potential of LAC12aGE in cancer models.

Main Methods:

• Development of a photo-responsive caged crRNA for CRISPR/Cas12a.
• Light-induced activation of the CRISPR/Cas12a machinery (LAC12aGE).
• Application of LAC12aGE for gene editing in cell lines and in vivo tumor models.

Main Results:

• LAC12aGE successfully edited exogenous and endogenous genes in living cells.
• Knockout of the hepatocyte growth factor (HGF) gene inhibited HepG2 cell proliferation and migration.
• In vivo administration of LAC12aGE effectively inhibited HepG2 tumor growth, inducing apoptosis and necrosis.

Conclusions:

• The photo-triggered CRISPR/Cas12a system provides precise spatiotemporal control over gene editing.
• LAC12aGE demonstrates significant potential for in vitro gene editing applications.
• LAC12aGE shows promise as a therapeutic strategy for inhibiting tumor growth in vivo.