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EPS and iPS Cells in Disease Research01:21

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Embryonic and induced pluripotent stem cells are excellent models for disease research because of their ability to self-renew and differentiate into most cell types. Somatic cells from a patient are isolated and reprogrammed into induced pluripotent stem cells or iPSCs. These iPSCs are later differentiated into the desired cell type, which mirrors the diseased cell of the patient. In this way, disease models have been created for investigating diseases such as Down syndrome, type I diabetes,...
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Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore...
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Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
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Related Experiment Video

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Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts
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Manipulating and studying gene function in human pluripotent stem cell models.

Elisa Balmas1, Federica Sozza1, Sveva Bottini1

  • 1Department of Molecular Biotechnology and Health Sciences, Molecular Biotechnology Center "Guido Tarone", University of Turin, Torino, Italy.

FEBS Letters
|July 31, 2023
PubMed
Summary
This summary is machine-generated.

This review details methods for manipulating gene function in human pluripotent stem cells (hPSCs). It covers gene editing tools like CRISPR/Cas9 and their applications in studying development and disease for regenerative medicine.

Keywords:
CRISPR interference and activationCRISPR/Cas9RNA interferencearrayed and pooled screensbase and prime editinggenomic safe harborshomologous recombinationhuman pluripotent stem cellssingle-cell screenstransgenesis

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Area of Science:

  • Stem Cell Biology
  • Genetics
  • Molecular Biology

Background:

  • Human pluripotent stem cells (hPSCs) are crucial for studying human development and disease.
  • Applications in regenerative medicine depend on effective gene function manipulation in hPSCs.

Purpose of the Study:

  • To provide a comprehensive review of gene manipulation techniques in hPSCs.
  • To guide researchers new to the field and update experienced stem cell biologists.

Main Methods:

  • Discusses challenges and solutions for gene expression manipulation (transfection, transduction, transposition, safe harbor editing).
  • Outlines historical (RNAi, transgenesis, homologous recombination) and modern (CRISPR/Cas9, base editing, prime editing) methods for loss-, gain-, and change-of-function studies.
  • Covers arrayed/pooled functional studies, single-cell genomics, and bioinformatic tools.

Main Results:

  • Highlights established and emerging technologies for precise gene editing in hPSCs.
  • Details strategies for constitutive or inducible gene modulation.
  • Explains the integration of these methods for functional genomic studies.

Conclusions:

  • Mastering these gene manipulation technologies will drive significant advancements in human biology and medicine.
  • The review empowers researchers to utilize advanced techniques for hPSC-based research.