Screening for novel factors involved in mouse early embryonic development using inhibitor libraries

Area of Science:

• Developmental Biology
• Genetics
• Pharmacology

Background:

• Mammalian early embryonic development involves numerous factors, many yet to be identified.
• Omics approaches and inhibitor screening are key to discovering these factors.

Purpose of the Study:

• To develop a novel screening system for identifying essential factors in mouse early embryonic development.
• To discover novel regulators of fertilized egg development using this system.

Main Methods:

• Developed a screening system combining ultra-superovulation and one-cell stage embryo cryopreservation in mice.
• Screened 95 inhibitors to identify factors crucial for mouse fertilized egg development.
• Utilized genome editing (gene knockout) to verify identified factors.

Main Results:

• Identified 16 essential factors, including 5 known and 11 novel regulators.
• Confirmed ATPases arrest development at distinct stages.
• Discovered novel regulators: p53 activator (PRIMA-1), cathepsin D, CXCR2, SK2, and SK3 potassium channels.
• Gene knockout of cathepsin D and CXCR2 confirmed their role in embryonic development arrest.

Conclusions:

• The developed screening method effectively identifies novel factors in embryonic development.
• This approach can discover species-specific regulators of early development across different species and inhibitor libraries.