Jove
Visualize
Contact Us

Related Concept Videos

CRISPR01:59

CRISPR

50.8K
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...
50.8K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.0K
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...
17.0K
Homologous Recombination02:31

Homologous Recombination

50.5K
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...
50.5K
Epigenetic Regulation01:37

Epigenetic Regulation

3.0K
Epigenetic changes alter the physical structure of the DNA without changing the genetic sequence and often regulate whether genes are turned on or off. This regulation ensures that each cell produces only proteins necessary for its function. For example, proteins that promote bone growth are not produced in muscle cells. Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
X-chromosome...
3.0K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
6.0K

You might also read

Related Articles

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

Sort by
Same author

Multiplex epigenome editing of <i>MECP2</i> to rescue Rett syndrome neurons.

Science translational medicine·2023
Same author

Trifactorial classification system for osteotome sinus floor elevation based on an observational retrospective analysis of 926 implants followed up to 10 years.

Quintessence international (Berlin, Germany : 1985)·2015
See all related articles
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 Experiment Video

Updated: Jun 27, 2025

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
07:49

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

Published on: May 30, 2025

869

CRISPR/dCas9-Tet1-Mediated DNA Methylation Editing.

Junming Qian1, Shawn X Liu1

  • 1Department of Physiology and Cellular Biophysics, Columbia University Medical Center, Columbia University, New York, NY, USA.

Bio-Protocol
|April 30, 2024
PubMed
Summary

Scientists developed a targeted DNA methylation editing tool (dCas9-Tet1) to precisely demethylate specific genomic loci. This system enables studying epigenetic roles and fine-tuning gene expression without altering DNA sequences.

Keywords:
CRISPRDNA methylationEpigenome editingGene regulationdCas9-Tet1

More Related Videos

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
10:44

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

1.4K
Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models
13:47

Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models

Published on: March 29, 2019

9.6K

Related Experiment Videos

Last Updated: Jun 27, 2025

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
07:49

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

Published on: May 30, 2025

869
In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
10:44

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

1.4K
Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models
13:47

Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models

Published on: March 29, 2019

9.6K

Area of Science:

  • Epigenetics
  • Molecular Biology
  • Genomics

Background:

  • DNA methylation is a crucial epigenetic mechanism implicated in biological processes and human diseases.
  • Previous methods for altering DNA methylation were largely non-specific, hindering locus-specific functional studies.
  • Aberrant DNA methylation is linked to various human diseases, necessitating precise manipulation tools.

Purpose of the Study:

  • To develop a novel tool for targeted DNA demethylation at specific genomic loci.
  • To enable precise investigation of the functional significance of DNA methylation.
  • To create a scalable and efficient system for epigenetic manipulation in cell cultures.

Main Methods:

  • Fusion of the catalytic domain of Ten-eleven translocation dioxygenase 1 (Tet1) with a deactivated Cas9 (dCas9) protein.
  • Development of a dCas9-Tet1 system for sequence-specific DNA demethylation guided by single-guide RNA (sgRNA).
  • Establishment of a protocol for modular and scalable application in various cell cultures.

Main Results:

  • Demonstrated high efficiency and specificity in targeted DNA demethylation at specific genomic loci.
  • Enabled fine-tuning of gene expression by manipulating DNA methylation without altering the underlying DNA sequence.
  • Showcased the applicability of the dCas9-Tet1 system across diverse cell culture types.

Conclusions:

  • The dCas9-Tet1 system provides a powerful and versatile tool for precise epigenetic editing.
  • This technology facilitates the study of DNA methylation's role in biological processes and disease.
  • The system holds potential for future ex vivo and in vivo therapeutic applications.