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

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...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

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...

Experimental RNAi

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

You might also read

Related Articles

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

Sort by

Same author

Discovery of Tcf7 regulators with clonally-resolved CRISPR screens identifies Trim28 as a mediator of CD8 T cell differentiation in tumors.

bioRxiv : the preprint server for biology·2026

Same author

Biological aging and generational shifts in early-onset cancer risk.

Nature medicine·2026

Same author

Fibroblast growth factor receptor inhibition for succinate dehydrogenase-deficient gastrointestinal stromal tumors: a phase 2 trial.

Nature medicine·2026

Same author

CRISPR base editor screening identifies spectrum of MEN1 mutations impacting menin inhibitors in clinical trials.

Nature communications·2026

Same author

CD164 is an endolysomal host factor for entry of Clade A New World Arenaviruses.

bioRxiv : the preprint server for biology·2026

Same author

Transcription factor collaboration enables precise T cell state engineering.

bioRxiv : the preprint server for biology·2026

Same journal

Kat5 deficiency in alveolar type II cells licenses STAT6-driven glycolytic reprogramming and pulmonary fibrosis.

Nature communications·2026

Same journal

Continuous nonthermal slab gap formed by progressive tearing beneath Northeast Asia.

Nature communications·2026

Same journal

Zeolitic isolated protonic acid sites-mediated NH<sub>3</sub> storage for robust NO<sub>x</sub> removal.

Nature communications·2026

Same journal

Coaxially nested component with asymmetric fiber resonant cavity and separation membrane for gaseous and dissolved gases detection.

Nature communications·2026

Same journal

Near-unity charge readout signal in a nonlinear resonator without matching the sensor dissipation.

Nature communications·2026

Same journal

Prokaryotic Schlafen proteins cleave tRNAs during type III CRISPR immunity.

Nature communications·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 29, 2025

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

Published on: April 25, 2022

Multi-locus CRISPRi targeting with a single truncated guide RNA.

Molly M Moore¹, Siddarth Wekhande¹, Robbyn Issner¹

¹Gene Regulation Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Nature Communications

|February 5, 2025

Summary

This summary is machine-generated.

Researchers developed a CRISPR interference (CRISPRi) method using truncated guides to efficiently screen thousands of potential transcription factor binding sites. This approach aids in understanding gene regulation and its role in cellular function and disease.

More Related Videos

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

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

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 29, 2025

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

Published on: April 25, 2022

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

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

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

Published on: June 16, 2017

Area of Science:

Genomics
Molecular Biology
Epigenetics

Background:

Functional genomics aims to identify non-coding elements influencing gene expression, cellular processes, and disease.
Characterizing these elements is difficult due to their vast number and complexity.

Purpose of the Study:

To develop a CRISPR interference (CRISPRi) screening method for efficiently evaluating numerous putative transcription factor binding sites.
To expand the targeting range of CRISPRi while preserving its gene-silencing capabilities.

Main Methods:

Developed a CRISPRi approach utilizing truncated guide RNAs (gRNAs) with a short spacer length (10 nucleotides).
Screened over 13,000 potential CTCF binding sites using 24 distinct truncated gRNAs.
Assessed the ability of truncated gRNAs to mediate the deposition of repressive H3K9me3 marks and disrupt transcription factor binding.

Main Results:

Truncated gRNAs effectively disrupted enhancer activity across multiple target sites.
CRISPRi-mediated deposition of H3K9me3 marks was observed at targeted loci.
Transcription factor binding was disrupted at a majority of sequence-matched target sites.

Conclusions:

The developed CRISPRi approach with truncated guides enables large-scale screening of transcription factor binding sites and other repetitive genomic elements.
This method offers a powerful and easily implementable tool for functional genomics research.
Facilitates the study of gene regulation and its implications in disease.