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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

607
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

CRISPR

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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

CRISPR and crRNAs

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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Related Experiment Video

Updated: Oct 12, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

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An Episomal CRISPR/Cas12a System for Mediating Efficient Gene Editing.

Nannan Duan1, Shuqing Tang1, Baitao Zeng1

  • 1Center for Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China.

Life (Basel, Switzerland)
|November 27, 2021
PubMed
Summary

A new CRISPR-based gene editing system, COE, significantly improves editing efficiency in stem cells. This breakthrough offers a safer, more effective tool for regenerative medicine applications.

Keywords:
CRISPR/Cas12aDMDOrip/EBNA1gene editingiPSCs

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Last Updated: Oct 12, 2025

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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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Area of Science:

  • Biomedical Science
  • Gene Editing Technology
  • Stem Cell Research

Background:

  • CRISPR technology offers powerful biomedical applications.
  • Low gene editing efficiency in stem cells limits regenerative medicine.
  • Induced pluripotent stem cells (iPSCs) require efficient editing tools.

Purpose of the Study:

  • To develop a novel gene editing system with enhanced efficiency.
  • To evaluate the safety and efficacy of the new system in stem cells.
  • To overcome limitations of current gene editing technologies in regenerative medicine.

Main Methods:

  • Designed and constructed the CRISPR/Cas12a and Orip/EBNA1-based COE gene-editing system.
  • Evaluated editing efficiency in HEK-293T cells using flow cytometry and RFLP analysis.
  • Verified COE efficiency in iPSCs via nucleofection, PCR, and Sanger sequencing.

Main Results:

  • COE achieved up to 90% insertion/deletion rates in HEK-293T cells.
  • Successfully deleted a 2.5 kb fragment (Exon 51) in iPSCs with high efficiency (3/14 positive clones).
  • Exon 51-deleted iPSCs showed similar expression profiles to DMD patient-specific iPSCs, with no plasmid residue.

Conclusions:

  • Developed a novel, efficient, and safe gene-editing system named COE.
  • COE provides a powerful new tool for advancing gene editing applications.
  • The system shows promise for regenerative medicine and studying genetic disorders like DMD.