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

Conservation of Protein Domains Over Different Proteins

Conservation of Protein Domains Over Different Proteins

Protein domains are small structurally independent units that are part of a single amino acid chain. Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...

Conservation of Protein Domains

Conservation of Protein Domains

Restarting Stalled Replication Forks

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...

Conserved Binding Sites

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...

Bacterial RNA Polymerase

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...

Protein Folding Quality Check in the RER

Protein Folding Quality Check in the RER

ER is the primary site for the maturation and folding of soluble and transmembrane secretory proteins. The calnexin cycle is a specific chaperone system that folds and assesses the confirmation of N-glycosylated proteins before they can exit the ER lumen. The primary players of this quality check pipeline are the lectins, ER-resident chaperones, and a glucosyl transferase enzyme. In case the calnexin system in the lumen fails to salvage a misfolded protein, it is transported to the cytoplasm...

You might also read

Related Articles

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

Sort by

Same author

Ultramutagenesis through combined Msh2 depletion and proofreading-deficient DNA polymerase ε expression enhances small molecule target identification in forward genetic screens.

bioRxiv : the preprint server for biology·2026

Same author

Structural and kinetic insights into a metagenomics-derived Cas12a with high specificity.

bioRxiv : the preprint server for biology·2026

Same author

Phage-encoded factor stimulates DNA degradation by the Hna anti-phage defense system.

Nature communications·2026

Same author

Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.

Nature reviews. Molecular cell biology·2026

Same author

Advancing Reproducibility and Open Data in Theoretical and Computational Chemistry.

Journal of chemical theory and computation·2026

Same author

Orthosteric and allosteric effects of anti-CRISPR II-C1 inhibition on <i>Geo</i> Cas9 from integrated structural biophysics.

bioRxiv : the preprint server for biology·2026

Search research articles

Related Experiment Video

Updated: Jun 22, 2025

Assessment of Immunologically Relevant Dynamic Tertiary Structural Features of the HIV-1 V3 Loop Crown R2 Sequence by ab initio Folding

Assessment of Immunologically Relevant Dynamic Tertiary Structural Features of the HIV-1 V3 Loop Crown R2 Sequence by ab initio Folding

Published on: September 15, 2010

Cas12a domain flexibility guides R-loop formation and forces RuvC resetting.

Isabel Strohkendl¹, Aakash Saha², Catherine Moy¹

¹Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712, USA.

|July 2, 2024

Summary

CRISPR-Cas12a

Keywords:

CRISPR-Cas Cas12a R-loop RNA-guided nuclease RuvC-mediated cleavage gene editing

More Related Videos

Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles

Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles

Published on: May 8, 2015

Design and Synthesis of a Reconfigurable DNA Accordion Rack

Design and Synthesis of a Reconfigurable DNA Accordion Rack

Published on: August 15, 2018

Related Experiment Videos

Last Updated: Jun 22, 2025

Assessment of Immunologically Relevant Dynamic Tertiary Structural Features of the HIV-1 V3 Loop Crown R2 Sequence by ab initio Folding

Assessment of Immunologically Relevant Dynamic Tertiary Structural Features of the HIV-1 V3 Loop Crown R2 Sequence by ab initio Folding

Published on: September 15, 2010

Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles

Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles

Published on: May 8, 2015

Design and Synthesis of a Reconfigurable DNA Accordion Rack

Design and Synthesis of a Reconfigurable DNA Accordion Rack

Published on: August 15, 2018

Area of Science:

Molecular Biology
Structural Biology
Biotechnology

Background:

CRISPR-Cas12a is an RNA-guided endonuclease with significant potential in biotechnology and therapeutics.
Understanding its mechanism is crucial for optimizing its applications.

Purpose of the Study:

To elucidate the structural mechanisms of CRISPR-Cas12a's DNA targeting and nuclease activation.
To detail the R-loop formation process and its role in target recognition.

Main Methods:

Cryo-electron microscopy (cryo-EM) was employed to visualize Cas12a R-loop intermediates.
Structural analysis captured dynamic stages of DNA binding and cleavage.

Main Results:

Distinct REC domain arrangements correlate with R-loop formation stages.
Domain flexibility facilitates non-target strand engagement and subsequent cleavage.
Target strand repositioning and RuvC active site occlusion/resetting were observed.

Conclusions:

A structural model for Cas12a DNA targeting, R-loop formation, and cleavage was established.
The findings provide mechanistic insights into Cas12a specificity and compare it to other Class 2 CRISPR effectors.