Mapping transcription factor functions in astrocytes using in vivo gain-of-function Perturb-seq

Area of Science:

• Neuroscience
• Genetics
• Systems Biology

Background:

• Understanding genotype-phenotype relationships in complex tissues requires advanced in vivo methods.
• Astrocytes play crucial roles in brain function, making them a key cell type for study.
• High-throughput screening combined with cell type-specific readouts is needed for functional genomics.

Purpose of the Study:

• To develop and apply an in vivo gain-of-function Perturb-seq platform (iGOF-Perturb-seq) for functional genomics in astrocytes.
• To create a functional atlas of approximately 1000 transcription factors (TFs) in astrocytes.
• To identify cofunctional TF modules, annotate unknown TFs, and predict disease-associated TF clusters.

Main Methods:

• Development of the iGOF-Perturb-seq platform for in vivo gain-of-function screening.
• Application of the platform to profile ~1000 transcription factors in mouse astrocytes.
• Utilized a mouse neuroinflammatory model to test therapeutic candidates.

Main Results:

• Generated a functional atlas of ~1000 transcription factors in astrocytes.
• Identified cofunctional TF modules and annotated uncharacterized TFs.
• Discovered Ferd3l as a therapeutic candidate; its astrocyte-specific overexpression alleviated Alzheimer's disease symptoms in mice.

Conclusions:

• The iGOF-Perturb-seq platform enables in vivo functional genomics in specific cell types.
• This study provides valuable resources for understanding gene regulation and brain disease mechanisms.
• Identified Ferd3l as a potential therapeutic target for Alzheimer's disease and other brain disorders.