A human tissue screen identifies a regulator of ER secretion as a brain-size determinant.
Christopher EskDominik LindenhoferSimon HaendelerRoelof A WesterFlorian G PflugBenoit SchroederJoshua A BagleyUlrich EllingJohannes ZuberArndt von HaeselerJuergen A KnoblichPublished in: Science (New York, N.Y.) (2020)
Loss-of-function (LOF) screens provide a powerful approach to identify regulators in biological processes. Pioneered in laboratory animals, LOF screens of human genes are currently restricted to two-dimensional cell cultures, which hinders the testing of gene functions requiring tissue context. Here, we present CRISPR-lineage tracing at cellular resolution in heterogeneous tissue (CRISPR-LICHT), which enables parallel LOF studies in human cerebral organoid tissue. We used CRISPR-LICHT to test 173 microcephaly candidate genes, revealing 25 to be involved in known and uncharacterized microcephaly-associated pathways. We characterized IER3IP1, which regulates the endoplasmic reticulum (ER) function and extracellular matrix protein secretion crucial for tissue integrity, the dysregulation of which results in microcephaly. Our human tissue screening technology identifies microcephaly genes and mechanisms involved in brain-size control.
Keyphrases
- genome wide
- endothelial cells
- zika virus
- endoplasmic reticulum
- dna methylation
- extracellular matrix
- intellectual disability
- crispr cas
- induced pluripotent stem cells
- pluripotent stem cells
- high throughput
- genome editing
- transcription factor
- stem cells
- multiple sclerosis
- white matter
- blood brain barrier
- small molecule
- single molecule
- subarachnoid hemorrhage
- estrogen receptor
- amino acid
- binding protein
- cerebral ischemia
- functional connectivity