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  11. Generation Of A Leutx-2a-mcherry Knock-in H1 Human Embryonic Stem Cell Line Using Crispr/cas9 System.
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  11. Generation Of A Leutx-2a-mcherry Knock-in H1 Human Embryonic Stem Cell Line Using Crispr/cas9 System.

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Generation of a LEUTX-2A-mCherry knock-in H1 human embryonic stem cell line using CRISPR/Cas9 system.

Xiu Yu1, Zezhong Su2

  • 1School of Medicine, Jiaying University, Meizhou 514015, China; Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China; Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-sen University, Guangzhou 510080, China.

Stem Cell Research
|June 13, 2025

View abstract on PubMed

Summary
This summary is machine-generated.

Researchers developed a new tool to track LEUTX expression, improving the study of early human development. This helps identify and analyze key cells for understanding the transition from pluripotency to totipotency.

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Area of Science:

  • Developmental Biology
  • Stem Cell Biology
  • Genomics

Background:

  • LEUTX is crucial for human embryonic genome activation at the 8-cell stage.
  • Induced 8-cell-like cells (8CLCs) model the pluripotency-to-totipotency transition.
  • Current 8CLC induction methods suffer from low efficiency and heterogeneity.

Purpose of the Study:

  • To develop a reporter system for dynamic monitoring of endogenous LEUTX expression.
  • To enable purification and functional characterization of genuine 8CLCs.
  • To facilitate the study of regulatory networks in totipotency acquisition.

Main Methods:

  • Development of a fluorescent reporter system for LEUTX.
  • Utilizing the reporter for purification of 8CLCs.
  • Functional characterization of purified 8CLCs.

    • Main Results:

    • The reporter system accurately reflects endogenous LEUTX expression dynamics.
    • Facilitated purification of a homogeneous population of bona fide 8CLCs.
    • Enabled detailed analysis of molecular mechanisms driving totipotency.

    Conclusions:

    • The LEUTX reporter system overcomes limitations in 8CLC induction.
    • Provides a powerful tool for dissecting early human development and totipotency.
    • Advances the study of stem cell transitions and genome activation.