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Related Concept Videos

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Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for...
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Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells
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Microcell-mediated chromosome transfer between non-identical human iPSCs.

Narumi Uno1,2, Hitomaru Miyamoto2, Kyotaro Yamazaki2,3,4

  • 1Laboratory of Bioengineering, Faculty of Life Sciences, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.

Molecular Therapy. Nucleic Acids
|December 5, 2024
PubMed
Summary

This study introduces a novel microcell-mediated chromosome transfer (MMCT) method using human induced pluripotent stem cells (hiPSCs) for creating disease models. This technique efficiently generates hyperaneuploidy in hiPSCs, aiding rare disease research.

Keywords:
Klinefelter’s syndromeMT: Delivery Strategiesdisease modelhuman artificial chromosomeinduced pluripotent stem cellmicrocell-mediated chromosome transfermouse artificial chromosomepaclitaxelreversinetriple X syndrometrisomy

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Area of Science:

  • Stem cell biology
  • Genetics
  • Developmental biology

Background:

  • Microcell-mediated chromosome transfer (MMCT) is a key technique for generating aneuploidy models.
  • Existing methods using mouse/hamster cell lines are prone to chromosomal rearrangements and are labor-intensive.
  • Human induced pluripotent stem cells (hiPSCs) offer a promising alternative for chromosome transfer and disease modeling.

Purpose of the Study:

  • To develop an efficient MMCT method utilizing hiPSCs as both donor and recipient cells.
  • To establish a robust system for generating hiPSC-based models of hyperaneuploidy syndromes.
  • To overcome limitations of traditional MMCT methods regarding chromosomal stability and generation time.

Main Methods:

  • Developed an MMCT protocol using hiPSCs, employing paclitaxel and reversine for micronucleation.
  • Utilized ecotropic viral envelope and mCAT-1 interactions for efficient membrane fusion during MMCT.
  • Employed CRISPR-Cas9 for tagging specific chromosomes (Chr21, ChrY) and transferring human/mouse artificial chromosomes (Chr21, ChrX, ChrY).

Main Results:

  • Successfully established an MMCT method using hiPSCs as donor and recipient cells.
  • Generated isogenic hiPSC disease models with controlled hyperaneuploidy.
  • Achieved full-length introduction of artificial chromosomes, a feat not previously reported.

Conclusions:

  • The developed CRISPR-Cas9 and MMCT strategy provides a systematic approach to create hiPSC disease models with hyperaneuploidy.
  • This method facilitates the study of rare genetic disorders and offers insights into early developmental mechanisms.
  • Enables the introduction of comprehensive chromosomal sets into hiPSCs for advanced research.