Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Animal Mitochondrial Genetics02:59

Animal Mitochondrial Genetics

7.4K
Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
7.4K
In-vitro Mutagenesis01:16

In-vitro Mutagenesis

13.7K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
13.7K
RNA Editing02:23

RNA Editing

8.9K
RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
8.9K
CRISPR01:59

CRISPR

49.2K
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...
49.2K
Mismatch Repair01:20

Mismatch Repair

4.8K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
4.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same authorSame journal

PTEN acts as a master mediator of nonhealing venous leg ulcers by suppressing immune response, angiogenesis, and lymphangiogenesis.

Science translational medicine·2026
Same author

Dopaminergic neurons preferentially accumulate mtDNA rearrangements.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Plasma proteomic profiles of Alzheimer's disease and neurodegeneration in African cohorts.

Nature communications·2026
Same author

Persistence of large mtDNA rearrangements linked to premature aging in Pol γ exonuclease-deficient mice.

Nucleic acids research·2026
Same author

GABA signaling activation drives glioblastoma progression in female mice through myeloid-derived suppressor cells.

Nature cancer·2026
Same author

Efficient prime editing in vivo and in vitro using lipid nanoparticles.

Nature nanotechnology·2026

Related Experiment Video

Updated: May 30, 2025

Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein
05:48

Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein

Published on: March 16, 2022

2.6K

Correcting a pathogenic mitochondrial DNA mutation by base editing in mice.

Jose D Barrera-Paez1, Sandra R Bacman2, Till Balla3

  • 1Graduate Program in Human Genetics, University of Miami Miller School of Medicine, 1501 NW 10th Avenue (M-860), Miami, FL 33136, USA.

Science Translational Medicine
|January 29, 2025
PubMed
Summary

Mitochondrial gene therapy using a DddA-derived cytosine base editor (DdCBE) successfully restored mitochondrial transfer RNA (tRNA) alanine levels in mice. However, careful dose monitoring is crucial to prevent off-target mitochondrial DNA editing and adverse effects.

More Related Videos

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
07:24

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

1.4K
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
08:22

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Published on: March 12, 2018

14.8K

Related Experiment Videos

Last Updated: May 30, 2025

Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein
05:48

Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein

Published on: March 16, 2022

2.6K
Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
07:24

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

1.4K
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
08:22

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Published on: March 12, 2018

14.8K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Primary mitochondrial disorders often stem from mutations in mitochondrial DNA (mtDNA).
  • A specific mutation (m.5024C→T) in the mitochondrial transfer RNA alanine (mt-tRNAAla) gene destabilizes its structure, leading to dysfunction.

Purpose of the Study:

  • To evaluate the efficacy of a DddA-derived cytosine base editor (DdCBE) for correcting a pathological mtDNA mutation.
  • To assess the potential of DdCBE as a gene therapy for mitochondrial disorders by restoring mt-tRNAAla function in a mouse model.

Main Methods:

  • A DdCBE construct was designed to introduce a compensatory edit (m.5081G→A) in the mt-tRNAAla gene.
  • The DdCBE was tested in vitro in a mutant cell line and delivered in vivo via adeno-associated virus 9 (AAV9) in mice.
  • Mitochondrial tRNA levels, lactate concentrations, and off-target editing were analyzed post-treatment.

Main Results:

  • In vitro editing increased mt-tRNAAla levels by up to 78% in mutant cells.
  • In vivo, DdCBE restored mt-tRNAAla amounts in heart and skeletal muscle in a dose-dependent manner, decreasing elevated lactate levels.
  • High AAV9-DdCBE doses caused severe adverse effects due to extensive off-target mtDNA editing.

Conclusions:

  • DdCBE shows promise as a gene therapy tool for mitochondrial disorders by correcting specific mtDNA mutations.
  • Careful dose optimization of DdCBE is essential to balance therapeutic benefits and mitigate risks of off-target editing and toxicity.