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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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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: Apr 1, 2026

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Efficient Mitochondrial Genome Editing by CRISPR/Cas9.

Areum Jo1, Sangwoo Ham1, Gum Hwa Lee2

  • 1Division of Pharmacology, Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon, Gyeonggi-do 440-746, Republic of Korea.

Biomed Research International
|October 9, 2015
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Summary

CRISPR/Cas9 technology can now edit mitochondrial DNA (mtDNA). Researchers developed a specialized mitochondria-targeted Cas9 (mitoCas9) for precise mtDNA editing, enabling new therapeutic strategies.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system is a powerful tool for nuclear DNA editing.
  • Its application for mitochondrial DNA (mtDNA) editing remains largely unexplored.
  • Mitochondria play crucial roles in cellular energy production and are implicated in various diseases.

Purpose of the Study:

  • To investigate the feasibility of using CRISPR/Cas9 for editing mitochondrial DNA.
  • To develop a mitochondria-specific CRISPR/Cas9 system for targeted mtDNA modification.
  • To assess the impact of mtDNA editing on cellular processes and mitochondrial function.

Main Methods:

  • Delivery of FLAG-Cas9 and guide RNAs (gRNAs) targeting specific mtDNA loci (Cox1, Cox3).
  • Development and utilization of a mitochondria-targeted Cas9 (mitoCas9) for enhanced specificity.
  • Analysis of mtDNA cleavage, mtDNA levels, transcription, mitochondrial membrane potential, and cell growth.

Main Results:

  • FLAG-Cas9 demonstrated localization to mitochondria and cleavage of targeted mtDNA loci.
  • Mitochondrial protein homeostasis was disrupted following mtDNA cleavage.
  • MitoCas9 specifically targeted and cleaved mtDNA, leading to reduced mtDNA levels and transcription.
  • MitoCas9-induced mtDNA reduction disrupted mitochondrial membrane potential and inhibited cell growth.

Conclusions:

  • CRISPR/Cas9 technology is applicable for mitochondrial DNA editing.
  • The novel mitoCas9 system provides specific and efficient mtDNA editing without affecting genomic DNA.
  • MitoCas9 holds promise for therapeutic applications targeting mitochondrial genome-related diseases.