bioRxiv : the preprint server for biologyNov 24, 2025

Genome-wide CRISPR screen finds Clip2 as a new controller of bone cell development and shape

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jun 27, 2026

Abstract

Essence

A genome-wide CRISPRi screen pointed to Clip2 as a regulator of osteocyte maturation markers and dendrite morphology.

Evidence

In vitro functional genomic screening in the Ocy454 osteocyte-like cell line used surface CD61 as a maturation endpoint and found that Clip2 inhibition reduced CD61 expression, osteocyte-specific gene expression, and dendrite morphology.

Caveat

The evidence is limited to a cell-line assay using CD61 as a phenotypic marker, so bone-level function and therapeutic relevance were not shown.

Simplified

Osteocytes play critical roles in bone, making them attractive targets for therapeutics to improve bone mass and strength. The genes driving osteocyte maturation and function are not fully understood. Here we aimed to identify novel genes responsible for osteocyte differentiation and dendrite development by performing a genome-wide CRISPR-interference (CRISPRi) screen in the Ocy454 osteocyte-like cell line. We identify CD61 (integrin β3) as a marker of osteocyte maturation: surface CD61 expression increases during osteocyte maturation, and CD61cells express higher levels of osteocyte marker genes. We then developed a flow cytometry-based assay to quantify surface CD61 protein levels as a phenotypic endpoint for functional genomic screening. In a genome-wide screen, we identified Clip2, which encodes a microtubule binding protein, as one of dozens of genes necessary for CD61 expression. Clip2 inhibition decreased surface CD61 expression, reduced expression of osteocyte-specific genesand, and impaired dendrite morphology. Together, these results highlight the utility of surface CD61 as a marker of osteocyte maturity and identify a role of the microtubule cytoskeleton for osteocyte differentiation, form, and function. high Dmp1Sost in vitro

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Genome-wide CRISPR screen finds Clip2 as a new controller of bone cell development and shape

Abstract

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