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Related Concept Videos

Chromatin Modification in iPS Cells01:32

Chromatin Modification in iPS Cells

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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.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
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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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Related Experiment Video

Updated: Mar 23, 2026

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
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Genome Editing in Human Pluripotent Stem Cells.

Cory Smith1, Zhaohui Ye2, Linzhao Cheng1

  • 1Division of Hematology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; Stem Cell Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; Predoctoral Training Program in Human Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.

Cold Spring Harbor Protocols
|April 3, 2016
PubMed
Summary

CRISPR-Cas9 technology enables precise DNA editing in pluripotent stem cells (PSCs). This advancement allows for targeted gene correction and functional analysis, making complex genome engineering accessible for research and regenerative medicine.

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Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
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Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells

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Area of Science:

  • Cell Biology
  • Genetics
  • Biotechnology

Background:

  • Pluripotent stem cells (PSCs) are crucial for biological research and disease modeling due to their self-renewal and differentiation capabilities.
  • Precise DNA editing in PSCs is essential for correcting mutations and analyzing genetic variations for research and regenerative medicine.

Purpose of the Study:

  • To describe principles for designing single guide RNA (sgRNA) for targeted gene editing in human PSCs.
  • To outline strategies for disrupting, inserting, or replacing specific DNA sequences within human PSCs using genome editing tools.

Main Methods:

  • Utilizing designer nucleases, specifically the CRISPR-Cas9 system, to create targeted double-strand breaks (DSBs) in human PSC DNA.
  • Employing engineered single guide RNA (sgRNA) to direct the Cas9 nuclease to specific genomic loci for precise editing.

Main Results:

  • Demonstrated efficient and precise genome editing in human PSCs using the CRISPR-Cas9 system.
  • Enabled targeted gene disruption, insertion, and replacement with high accuracy and ease.

Conclusions:

  • CRISPR-Cas9 technology significantly enhances the ability to engineer human PSCs with unprecedented precision.
  • These advancements democratize sophisticated genome engineering techniques, previously confined to specialized facilities, for broader research applications.