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Related Concept Videos

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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CRISPR01:59

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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...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Related Experiment Video

Updated: Mar 8, 2026

CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions
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CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions

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Precise and efficient scarless genome editing in stem cells using CORRECT.

Dylan Kwart1, Dominik Paquet1, Shaun Teo1

  • 1Laboratory for Brain Development and Repair, The Rockefeller University, New York, USA.

Nature Protocols
|January 20, 2017
PubMed
Summary
This summary is machine-generated.

This study introduces CORRECT, a novel genome-editing framework that precisely modifies DNA using CRISPR/Cas9 technology. It overcomes previous limitations to accurately and efficiently create scarless genetic changes in stem cells.

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

Published on: September 25, 2019

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 enables precise DNA editing for disease modeling in stem cells.
  • Unwanted mutations (indels) arise from CRISPR/Cas9 re-editing, reducing accuracy.
  • Accurate genome editing is crucial for developing reliable in vitro disease models.

Purpose of the Study:

  • To develop a novel genome-editing framework to enhance CRISPR/Cas9 accuracy.
  • To enable scarless and efficient introduction of specific base changes in stem cells.
  • To provide a detailed protocol for implementing the CORRECT framework.

Main Methods:

  • Developed the CORRECT (consecutive re-guide or re-Cas steps to erase CRISPR/Cas-blocked targets) framework.
  • Utilized two rounds of genome editing with blocking mutations.
  • Applied the framework to human induced pluripotent stem cells (iPSCs).

Main Results:

  • Achieved accurate, efficient, and scarless introduction of specific base changes.
  • Demonstrated successful monoallelic and biallelic sequence modifications.
  • Generated isogenic stem cell lines with intended genetic alterations.

Conclusions:

  • The CORRECT framework significantly improves CRISPR/Cas9 editing accuracy.
  • This method facilitates the generation of precise genetic models for human diseases.
  • The protocol enables the creation of scarlessly edited stem cell lines within approximately 3 months.