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/Cas9 Genome Editing

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...

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...

CRISPR and crRNAs

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...

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...

Homologous Recombination

Homologous Recombination

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...

You might also read

Related Articles

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

Sort by

Same author

Enhanced Cas12i3 system enables precise OsAUX3 editing for rice grain improvement.

Journal of integrative plant biology·2026

Same author

Elevating RNA m<sup>5</sup>C methylation provides a promising strategy for crop productivity.

National science review·2026

Same author

Improving iSpyMacCas9 multiplex genome editing in rice by CRISPR-combo-mediated BBM1 activation.

The Plant journal : for cell and molecular biology·2026

Same author

Boosting genome editing of non-coding sequences in plants with glycosylase-mediated multi-nucleotide deletion editors.

Science bulletin·2026

Same author

A conserved boundary-specific NAC transcription factor targets various regulators to modulate rice aerial architecture.

The Plant cell·2026

Same author

Telomere-to-telomere genome assembly of Oryza australiensis reveals transposon-driven centromere repositioning and shared EE-DD ancestry.

Nature communications·2026

Search research articles

Related Experiment Video

Updated: Oct 16, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

CRISPR-BETS: a base-editing design tool for generating stop codons.

Yuechao Wu^1,2, Yao He³, Simon Sretenovic⁴

¹Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Jiangsu Key Laboratory of Crop Genetics and Physiology, Agricultural College of Yangzhou University, Yangzhou, China.

Plant Biotechnology Journal

|October 20, 2021

Summary

Cytosine base editors (CBEs) offer a precise method for creating gene knockouts by inserting stop codons. A new tool, CRISPR-BETS, simplifies designing guide RNAs for efficient plant gene knockout generation.

Keywords:

cytosine base editors guide RNA design plants protospacer adjacent motif stop codons

More Related Videos

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Published on: February 28, 2021

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: Oct 16, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Published on: February 28, 2021

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:

Molecular Biology
Genetics
Bioengineering

Background:

Cytosine base editors (CBEs) enable precise genome editing by introducing stop codons, facilitating genetic knockouts.
CBEs offer a predictable method for generating homozygous mutants efficiently without altering genome size.
Advancements in CBEs' activity, purity, and specificity have increased their appeal for gene knockout applications in plants and animals.

Purpose of the Study:

To address the lack of user-friendly design tools for employing CBEs to generate gene knockouts, particularly in plants.
To develop and validate a computational tool for designing guide RNAs (gRNAs) for CBE-mediated stop codon introduction.

Main Methods:

Development of CRISPR-BETS, a user-friendly design tool for gRNA design.
Application of CRISPR-BETS for designing gRNAs to introduce stop codons in protein-coding genes.
Experimental validation of CRISPR-BETS-designed gRNAs in rice and tomato.

Main Results:

CRISPR-BETS was demonstrated to be easy-to-use for gRNA design.
The tool generated highly specific and efficient gRNAs for introducing stop codons.
Successful generation of homozygous knockout lines in rice and tomato was achieved.

Conclusions:

CRISPR-BETS is an effective tool for designing gRNAs to generate gene knockouts using CBEs.
The tool simplifies the process of creating homozygous mutants in plants.
CRISPR-BETS is valuable for plant research and applicable to non-plant species.