Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Animal Mitochondrial Genetics

Animal Mitochondrial Genetics

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...

RNA Editing

RNA Editing

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...

CRISPR

CRISPR

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...

You might also read

Related Articles

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

Sort by

Same author

Efficient and reliable measles reprogramming platform for the generation of human iPSC.

Molecular therapy. Advances·2026

Same author

Herpes simplex virus alkaline nuclease interacts directly with ubiquitin-specific protease 15 through a conserved C-terminal domain.

The Journal of biological chemistry·2026

Same author

Validation and analysis of 12,000 AI-driven CAR-T designs in the <i>Bits to Binders</i> competitions.

bioRxiv : the preprint server for biology·2026

Same author

Rewiring STAT signaling from the cell surface with Trikine immunotherapeutics.

Science (New York, N.Y.)·2026

Same author

ercc6 deficient zebrafish exhibit UV and metronidazole sensitivity, increased oxygen consumption, and impaired hair cell mechanoelectrical transduction which can be restored by the superoxide dismutase mimetic MnTBAP.

Human molecular genetics·2026

Same author

Epigenome-wide association study of nuclear DNA methylation in relation to mitochondrial heteroplasmy.

Nature communications·2025

Same journal

Genome Editing for Familial Hemophagocytic Lymphohistiocytosis: Design Principles, Challenges, and Translational Perspectives.

Human gene therapy·2026

Same journal

Lilly, Ascidian Launch Up-to-$1.9B RNA Exon Editor Collaboration.

Human gene therapy·2026

Same journal

Hyperthermia-Enhanced Adeno-Associated Virus Gene Therapy: Mechanisms, Delivery Technologies, and Translational Potential.

Human gene therapy·2026

Same journal

Adeno-Associated Virus Gene Therapy Translation: Lessons from Early Regulatory Meetings.

Human gene therapy·2026

Same journal

In Remembrance of Professor Zelig Eshhar: <i>A Life Committed to CAR-T</i>.

Human gene therapy·2026

Same journal

<i>In Vivo</i> Genome Editing Approach to Disrupt Hydroxyacid Oxidase 1 for the Treatment of Primary Hyperoxaluria Type 1.

Human gene therapy·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jun 14, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Unconstrained Precision Mitochondrial Genome Editing with αDdCBEs.

Santiago R Castillo^1,2, Brandon W Simone³, Karl J Clark³

¹Virology and Gene Therapy Graduate Program, Mayo Clinic, Rochester, Minnesota, USA.

Human Gene Therapy

|August 30, 2024

Summary

This summary is machine-generated.

Engineered base editors (αDdCBEs) overcome the 5'-T constraint for mitochondrial DNA editing, enabling precise C•G-to-T•A conversions at previously inaccessible sites. This advancement expands therapeutic targets for mitochondrial genetic disorders.

Keywords:

DddA-derived cytosine base editors (DdCBEs)mitochondrial DNA (mtDNA)mitochondrial base editing precision genome engineering transcription activator-like effectors (TALEs)

More Related Videos

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

Published on: December 16, 2022

Related Experiment Videos

Last Updated: Jun 14, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

Published on: December 16, 2022

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

DddA-derived cytosine base editors (DdCBEs) are crucial for C•G-to-T•A conversions in mitochondrial DNA (mtDNA).
A 5'-T constraint limits DdCBE accessibility to over 150 human mtDNA loci.
Previous methods to bypass this constraint yielded suboptimal specificity.

Purpose of the Study:

To challenge the 5'-T constraint in DdCBE-mediated mtDNA editing.
To expand the range of editable motifs in mitochondrial DNA.
To develop more efficient and specific base editing technologies for mtDNA.

Main Methods:

Engineered DdCBEs (αDdCBEs) with TALE proteins recognizing all 5' bases were generated.
The activity and specificity of αDdCBEs were evaluated at various mtDNA loci.
Compatibility with DddA_tox variants (DddA6, DddA11) and TALE shifting was assessed.

Main Results:

αDdCBEs demonstrated efficient and specific mtDNA editing across diverse loci, irrespective of the 5' base.
αDdCBEs outperformed canonical DdCBEs in activity and specificity.
αDdCBEs are compatible with enhanced DddA_tox variants and TALE shifting for optimized editing.

Conclusions:

αDdCBEs enable unconstrained, efficient, and specific mitochondrial base editing.
This technology broadens the scope of potential therapeutic targets for mtDNA disorders.
αDdCBEs represent a significant advancement in mitochondrial genome engineering.