WITHDRAWN: High throughput CRISPR perturbation screens identify epigenetic regulators impacting primordial germ cell...

Area of Science:

• Epigenetics
• Developmental Biology
• Reproductive Science

Background:

• Environmental factors can negatively impact fertility and reproductive health.
• Epigenetic alterations in the germline are a potential mechanism for these effects.

Purpose of the Study:

• To identify genes and chemical compounds that perturb germline development using a large-scale CRISPRi screen.
• To investigate the role of epigenetic regulators in the differentiation of embryonic stem cells (ESCs) into primordial germ cell-like cells (PGCLCs).

Main Methods:

• Conducted a CRISPR interference (CRISPRi) screen of 701 epigenetic-related genes.
• Utilized a scalable, cytokine-free platform for ESC to PGCLC differentiation.
• Administered histone deacetylase inhibitors (HDACi) like valproic acid (VPA) and sodium butyrate (SB) to differentiating cells and mouse embryos.
• Performed transcriptome analysis on treated cells.

Main Results:

• Inhibition of 53 genes, notably NCOR2, significantly reduced PGCLC formation efficiency.
• HDAC inhibitors (VPA and SB) suppressed ESC to PGCLC differentiation.
• Exposure to VPA or SB in developing mouse embryos resulted in hypospermatogenesis.
• Transcriptome analysis revealed suppressed germline pathways and enhanced somatic pathways upon HDACi treatment.

Conclusions:

• NCOR2 and histone deacetylase activity are critical for proper germline differentiation.
• Environmental factors, including certain chemicals, can disrupt germline development and potentially affect fertility.
• Large-scale functional screens are effective for identifying agents impacting germline development.