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CRISPR loss of function screening to identify genes involved in human primordial germ cell-like cells development

Young Sun Hwang, M. Andrés Blanco, Kotaro Sasaki
doi: https://doi.org/10.1101/2022.05.23.493064
Young Sun Hwang
1Department of Biomedical Sciences, University of Pennsylvania, School of Veterinary Medicine, Philadelphia, PA 19104, USA
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M. Andrés Blanco
1Department of Biomedical Sciences, University of Pennsylvania, School of Veterinary Medicine, Philadelphia, PA 19104, USA
2Institute for Regenerative Medicine, University of Pennsylvania, PA 19104, USA
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  • For correspondence: ablanco@vet.upenn.edu ksasaki@upenn.edu
Kotaro Sasaki
1Department of Biomedical Sciences, University of Pennsylvania, School of Veterinary Medicine, Philadelphia, PA 19104, USA
2Institute for Regenerative Medicine, University of Pennsylvania, PA 19104, USA
3Department of Pathology and Laboratory Medicine, University of Pennsylvania, Perelman School of Medicine, Philadelphia, PA 19104, USA
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  • For correspondence: ablanco@vet.upenn.edu ksasaki@upenn.edu
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ABSTRACT

Despite our increasing knowledge of molecular mechanisms guiding various aspects of human reproduction, those underlying human primordial germ cell (PGC) development remain largely unknown. Here, we conducted custom CRISPR screening in an in vitro system of hPGC-like cells (hPGCLCs) to identify genes required for acquisition and maintenance of PGC fate in humans. Amongst our candidates, we identified TCL1A, an AKT coactivator. Functional assessment in our in vitro hPGCLCs system revealed that TCL1A played a critical role in later stages of hPGCLC development. Moreover, we found that TCL1A loss reduced AKT-mTOR signaling, downregulated expression of genes related to translational control, and subsequently led to a reduction in global protein synthesis and proliferation. Together, our study identifies novel regulators critical for hPGCLC development and demonstrates the importance of translational control in human reproduction.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted May 23, 2022.
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CRISPR loss of function screening to identify genes involved in human primordial germ cell-like cells development
Young Sun Hwang, M. Andrés Blanco, Kotaro Sasaki
bioRxiv 2022.05.23.493064; doi: https://doi.org/10.1101/2022.05.23.493064
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CRISPR loss of function screening to identify genes involved in human primordial germ cell-like cells development
Young Sun Hwang, M. Andrés Blanco, Kotaro Sasaki
bioRxiv 2022.05.23.493064; doi: https://doi.org/10.1101/2022.05.23.493064

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