Massively parallel in vivo Perturb-seq reveals cell-type-specific transcriptional networks in cortical development.
Cell
; 187(13): 3236-3248.e21, 2024 Jun 20.
Article
in En
| MEDLINE
| ID: mdl-38772369
ABSTRACT
Leveraging AAVs' versatile tropism and labeling capacity, we expanded the scale of in vivo CRISPR screening with single-cell transcriptomic phenotyping across embryonic to adult brains and peripheral nervous systems. Through extensive tests of 86 vectors across AAV serotypes combined with a transposon system, we substantially amplified labeling efficacy and accelerated in vivo gene delivery from weeks to days. Our proof-of-principle in utero screen identified the pleiotropic effects of Foxg1, highlighting its tight regulation of distinct networks essential for cell fate specification of Layer 6 corticothalamic neurons. Notably, our platform can label >6% of cerebral cells, surpassing the current state-of-the-art efficacy at <0.1% by lentivirus, to achieve analysis of over 30,000 cells in one experiment and enable massively parallel in vivo Perturb-seq. Compatible with various phenotypic measurements (single-cell or spatial multi-omics), it presents a flexible approach to interrogate gene function across cell types in vivo, translating gene variants to their causal function.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Gene Regulatory Networks
/
Single-Cell Analysis
Limits:
Animals
/
Female
/
Humans
Language:
En
Journal:
Cell
Year:
2024
Document type:
Article
Affiliation country:
United States