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Related Concept Videos

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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

Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing
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A Method for 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

Researchers can now easily edit human pluripotent stem cells (PSCs) to study genetic variants. This genome editing protocol allows precise mutations for disease modeling and regenerative medicine applications.

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

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Area of Science:

  • Stem cell biology
  • Genetics
  • Regenerative medicine

Background:

  • Human pluripotent stem cells (PSCs) are valuable for regenerative medicine, disease modeling, and basic research.
  • Precisely altering specific DNA sequences in PSCs is crucial for understanding genetic variant functions.
  • Existing methods require sophisticated systems for targeted genome editing in PSCs.

Purpose of the Study:

  • To describe a simplified protocol for genome editing in human pluripotent stem cells (PSCs).
  • To enable precise mutation induction (knockout, knock-in, single nucleotide alteration) in PSCs.
  • To facilitate the study of genetic variant effects and potential therapeutic applications.

Main Methods:

  • Detailed targeting schemes for genome editing in PSCs.
  • Protocols for electroporation, clonal isolation, and screening of edited cells.
  • Methods to induce specific mutations (knockout, knock-in, single nucleotide changes).

Main Results:

  • A simplified genome editing procedure for PSCs is presented.
  • The protocol allows for precise induction of desired mutations.
  • Researchers can now more easily generate isogenic PSC lines for comparative studies.

Conclusions:

  • Recent advances simplify genome editing in PSCs, making it accessible to individual researchers.
  • This protocol supports the study of gene function and the reversal of disease-causing variants.
  • Facilitates future applications in regenerative medicine and personalized therapies.