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Related Concept Videos

CRISPR01:59

CRISPR

52.7K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
52.7K
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Updated: Aug 18, 2025

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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CRISPR Manipulations in Stem Cell Lines.

Ya-Ju Chang1,2, Xuan Cui1,2, Sarah R Levi1,2

  • 1Edward S. Harkness Eye Institute, Department of Ophthalmology, Columbia University Irving Medical Center, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 9, 2022
PubMed
Summary
This summary is machine-generated.

Genome engineering using CRISPR-Cas9 technology enables the creation of precise cellular models, such as organoids and induced pluripotent stem cells (iPSCs), for disease research and drug discovery.

Keywords:
Gene editingHuman embryonic stem cellsHuman induced pluripotent stem cellsLociRibonucleoprotein

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Area of Science:

  • Cell biology
  • Genetics
  • Biotechnology

Background:

  • Organoids and patient-derived cells serve as valuable models for studying human diseases.
  • CRISPR-Cas9 technology has revolutionized genome engineering, enabling targeted gene modifications.
  • Induced pluripotent stem cells (iPSCs) offer a versatile platform for generating various cell lineages for research.

Purpose of the Study:

  • To detail advancements in genome engineering, focusing on CRISPR-Cas9 applications.
  • To describe the generation of targeted mutants in organoids and iPSCs for disease modeling.
  • To provide protocols for efficient gene editing in iPSCs using CRISPR-Cas9.

Main Methods:

  • Utilizing CRISPR-Cas9 nuclease system for targeted double-strand breaks in iPSCs.
  • Employing ribonucleoprotein (RNP) delivery for direct gene editing in cells.
  • Developing protocols for reagent preparation, transfection, and genotyping of iPSC clones.

Main Results:

  • High efficiency in gene editing of target loci in iPSCs achieved.
  • Demonstrated capability to generate targeted mutants in organoids and iPSCs.
  • Established protocols for RNP delivery and iPSC clone genotyping.

Conclusions:

  • CRISPR-Cas9 technology significantly advances cell engineering for basic research and therapeutics.
  • Modified organoids and iPSCs are powerful tools for disease modeling and drug testing.
  • Efficient gene editing protocols facilitate the study of gene function and disease mechanisms.