Low-input CUT&Tag for efficient epigenomic profiling of zebrafish stage I oocytes

Area of Science:

• Developmental Biology
• Epigenetics
• Molecular Biology

Background:

• Histone modifications are crucial epigenetic marks regulating gene expression.
• Oocyte epigenetic modifications significantly influence early embryonic development.
• Profiling oocyte epigenetics is challenging due to surrounding somatic cells.

Purpose of the Study:

• To establish a low-input CUT&Tag method for zebrafish stage I oocytes.
• To enable high-resolution profiling of histone modifications and DNA-binding proteins in oocytes.
• To investigate epigenetic regulation in oocyte development and maternal contribution.

Main Methods:

• Developed and optimized a low-input CUT&Tag protocol for zebrafish stage I oocytes.
• Established a workflow for isolating pure oocytes, library construction, and quality control.
• Applied CUT&Tag to profile histone modifications in wild-type and mutant zebrafish oocytes.

Main Results:

• Successfully profiled distinct histone modification patterns in zebrafish oocytes.
• Identified differentially expressed genes in oocytes with and without granulosa cells.
• Detected divergent histone modifications in wild-type versus *huluwa* mutant oocytes.

Conclusions:

• The low-input CUT&Tag method overcomes technical hurdles in zebrafish oocyte epigenetics.
• This technique provides a foundation for studying epigenetic mechanisms of maternal contribution.
• Enables deeper understanding of chromatin dynamics during oogenesis.