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In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
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A Novel Reporter Mouse Uncovers Endogenous Brn3b Expression
Adam M Miltner1, Yesica Mercado-Ayon2, Simranjeet K Cheema3
1Department of Cell Biology and Human Anatomy, University of California-Davis, Davis, CA 95616, USA. ammiltner@ucdavis.edu.
International Journal of Molecular Sciences
|June 15, 2019
Summary
We developed a Brn3b-mCherry reporter mouse using CRISPR technology. This tool allows noninvasive live imaging of retinal ganglion cells (RGCs) for monitoring visual system development and disease.
Area of Science:
- Neuroscience
- Developmental Biology
- Genetics
Background:
- Brn3b (Pou4f2) is a crucial transcription factor for neuronal development in the central nervous system.
- Retinal ganglion cells (RGCs) are vital for transmitting visual information from the retina to the brain.
Purpose of the Study:
- To create a Brn3b-mCherry reporter mouse line to visualize Brn3b expression.
- To establish a noninvasive method for longitudinal monitoring of RGCs.
Main Methods:
- CRISPR-based genetic engineering was employed to generate the Brn3b-mCherry reporter mouse.
- Expression patterns of Brn3b-mCherry were analyzed in various neural tissues.
- Scanning Laser Ophthalmoscopy (SLO) was used for live imaging of RGCs.
Main Results:
- The Brn3b-mCherry reporter accurately reflects endogenous Brn3b expression in the retina, optic tracts, midbrain tectum, and trigeminal ganglia.
- Novel Brn3b expression was detected in early optic stalk neuroectodermal cells.
- High fluorescent intensity enabled noninvasive live imaging of RGCs via SLO.
Conclusions:
- The Brn3b-mCherry reporter mouse provides a faithful readout of Brn3b expression during development.
- This reporter line offers a novel, noninvasive tool for in vivo, longitudinal studies of RGCs and visual system development.
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