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

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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...
CRISPR01:59

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 Short...
CRISPR01:59

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 Short...
CRISPR and crRNAs02:53

CRISPR and crRNAs

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...

You might also read

Related Articles

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

Sort by
Same author

Efficient genome editing with chimeric oligonucleotide-directed editing.

Nature communications·2026
Same author

Architecture of a DNA-guided Cas12a.

bioRxiv : the preprint server for biology·2026
Same author

Exploring the temperature stability of CRISPR-Cas12b using molecular dynamics simulations.

Molecular systems design & engineering·2025
Same author

AsCas12a tolerates insertions in target DNA.

Nucleic acids research·2025
Same author

DNA-guided CRISPR/Cas12 for RNA targeting.

Research square·2025
Same author

DNA-guided CRISPR/Cas12 for RNA targeting.

medRxiv : the preprint server for health sciences·2024

Related Experiment Video

Updated: May 17, 2026

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

DNA-guided CRISPR-Cas12 for cellular RNA targeting.

Carlos Orosco1, Boyu Huang1, Santosh R Rananaware1,2

  • 1Department of Chemical Engineering, University of Florida, Gainesville, FL, USA.

Nature Biotechnology
|May 15, 2026
PubMed
Summary

Researchers developed ΨDNA, a novel DNA guide for Cas12 nucleases to target RNA, enabling sensitive RNA detection and cellular RNA knockdown. This adaptable toolkit expands Cas12 applications beyond genome editing.

More Related Videos

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
08:37

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
11:35

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Related Experiment Videos

Last Updated: May 17, 2026

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
08:37

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
11:35

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • CRISPR Technology

Background:

  • Traditional CRISPR systems rely on RNA-guided nucleases for DNA or RNA targeting.
  • Cas12 nucleases are versatile tools for genome editing and diagnostics.
  • Developing novel guide systems can expand the capabilities of Cas12 effectors.

Purpose of the Study:

  • To engineer a DNA-based guide (ΨDNA) for RNA targeting by Cas12 nucleases.
  • To demonstrate the utility of ΨDNA for sensitive RNA detection and cellular RNA knockdown.
  • To explore the potential of ΨDNA for simultaneous DNA editing and RNA modulation.

Main Methods:

  • Engineering ΨDNA to mimic a reverse-oriented crRNA scaffold.
  • Utilizing AsCas12a and Cas12i1 nucleases for RNA recognition and trans-cleavage.
  • Assessing RNA detection sensitivity in clinical samples and multiplex RNA knockdown in cell lines.
  • Investigating the mechanism of ΨDNA activity through mechanistic studies.
  • Exploring modular fusions of Cas12a with other enzymes for expanded functions.

Main Results:

  • ΨDNA successfully guides Cas12 nucleases for RNA targeting, overcoming RNA-guided limitations.
  • Achieved 100% accurate hepatitis C virus RNA detection in clinical samples.
  • Demonstrated 70-95% multiplex knockdown of endogenous RNA transcripts via ribosome stalling.
  • Identified a critical stem loop structure for Cas12-ΨDNA-RNA complex stabilization.
  • Enabled simultaneous DNA editing and RNA knockdown, and expanded to RNA degradation and epitranscriptomic editing.

Conclusions:

  • ΨDNA guides offer a versatile platform for RNA targeting with Cas12 nucleases.
  • This technology enables precise and programmable control over cellular transcriptomes.
  • ΨDNA expands the Cas12 toolkit for applications in diagnostics, RNA regulation, and epitranscriptomics.