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Parallel functional testing identifies enhancers active in early postnatal mouse brain.

Jason T Lambert^1,2, Linda Su-Feher^1,2, Karol Cichewicz^1,2

¹Department of Psychiatry and Behavioral Sciences, University of California, Davis, Davis, United States.

|October 4, 2021

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This summary is machine-generated.

Researchers adapted massively parallel reporter assays (MPRAs) to screen for brain enhancers in mice. They identified novel enhancers, including those near CACNA1C, linked to neuropsychiatric disorders.

Area of Science:

Neuroscience
Genetics
Molecular Biology

Background:

Enhancers are cis-regulatory elements crucial for gene expression in the brain.
Massively parallel reporter assays (MPRAs) allow high-throughput screening of DNA sequences for enhancer activity.
Understanding non-coding sequences' role in development and disease is vital.

Purpose of the Study:

To adapt MPRA for in vivo screening of enhancer function in the early postnatal mouse brain.
To identify and validate novel brain-specific enhancers.
To investigate regulatory elements associated with neuropsychiatric disorders.

Main Methods:

Adapted a self-transcribing regulatory element MPRA strategy.
Delivered the MPRA system to the mouse brain using recombinant adeno-associated virus (rAAV).

Keywords:

MPRA enhancer genetics genomics mouse neurodevelopment neuroscience

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Performed paired screening and single enhancer in vivo functional testing.

Main Results:

Successfully identified and validated putative enhancers driving reporter gene expression in the mouse forebrain.
Discovered regulatory elements within an intronic CACNA1C linkage disequilibrium block.
Demonstrated the utility of rAAV-mediated MPRA for in vivo enhancer characterization.

Conclusions:

rAAV-delivered MPRA is a powerful tool for large-scale in vivo enhancer screening in the brain.
Identified novel enhancers, including those linked to neuropsychiatric disorder risk genes like CACNA1C.
This approach advances the understanding of enhancer function in brain development and disease.