A noninvasive photoactivatable split-Cre recombinase system for genome engineering in zebrafish

Area of Science:

• Developmental Biology
• Genetics and Genomics
• Molecular Biology

Background:

• The Cre/loxP system is widely used for in vivo cell labeling and tracking.
• Current limitations include challenges in achieving high spatiotemporal precision due to promoter reliance.
• Light-inducible systems offer advantages in regional confinement, tunability, and non-invasiveness for lineage tracing.

Purpose of the Study:

• To develop an efficient, genetically encoded, light-inducible transgenic Cre/loxP system in zebrafish.
• To overcome the limitations of conventional tissue-specific promoters for precise cell tracking.
• To enable non-invasive, spatiotemporally controlled cell fate and migration studies in vivo.

Main Methods:

• Development of a novel magnets-based, light-inducible transgenic Cre/loxP system in zebrafish.
• Utilized blue light illumination for non-invasive activation of the system.
• Assessed system performance for phototoxicity, leakiness, and recombination efficiency across various tissues and developmental stages.

Main Results:

• The developed system demonstrated no phototoxicity or leakiness in the dark.
• Efficient and robust Cre/loxP recombination was achieved in diverse tissues and cell types.
• Recombination occurred across different developmental stages upon non-invasive blue light exposure.

Conclusions:

• A novel, easy-to-use, light-inducible transgenic Cre/loxP system was successfully established in zebrafish.
• This system provides precise spatiotemporal control for in vivo cell tracking and lineage analysis.
• The tool is expected to advance research in light-controlled cell fate and migration studies.