Mouse primordial germ-cell-like cells lack piRNAs
Navin B Ramakrishna1, Giorgia Battistoni2, M Azim Surani3
1Wellcome/CRUK Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK; Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK; Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EL, UK.
Developmental Cell
|December 6, 2022
Summary
Day 6 mouse primordial germ cell-like cells (mPGCLCs) differentiated in vitro do not express piRNA pathway factors or detect piRNAs. These cells do not yet model later-stage mouse gonocytes where the piRNA pathway is active.
Area of Science:
- Reproductive biology
- Epigenetics
- RNA biology
Background:
- PIWI-interacting RNAs (piRNAs) are crucial for silencing transposable elements.
- Fetal piRNAs emerge in mouse gonocytes around embryonic day 13.5.
- In vitro differentiation of mouse primordial germ cell-like cells (mPGCLCs) offers a model to study the piRNA pathway.
Purpose of the Study:
- To investigate the expression of piRNA pathway factors and the presence of piRNAs during in vitro mPGCLC differentiation.
- To determine if Day 6 mPGCLCs accurately model the fetal piRNA pathway observed in vivo.
Main Methods:
- Single-cell RNA-seq and RT-qPCR were used to assess piRNA pathway factor expression.
- Small RNA sequencing was employed to detect piRNAs in mPGCLC lines.
Main Results:
- PiRNA pathway factors were not fully expressed in Day 6 mPGCLCs.
- No piRNAs were detected in a panel of Day 6 mPGCLC lines.
- Day 6 mPGCLCs do not resemble E13.5 or later mouse gonocytes regarding piRNA pathway activity.
Conclusions:
- In vitro differentiated Day 6 mPGCLCs are not yet suitable models for studying the active fetal piRNA pathway.
- Further optimization or different time points are needed for in vitro studies of the fetal piRNA pathway in mouse germ cells.
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