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

211
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...
211
CRISPR01:59

CRISPR

52.9K
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...
52.9K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.3K
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...
17.3K
Homologous Recombination02:31

Homologous Recombination

51.5K
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...
51.5K

You might also read

Related Articles

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

Sort by
Same author

The MAPK/ATF3/ASNS axis drives amino acid metabolic reprogramming to promote NSCLC survival under glucose deprivation.

Journal of translational medicine·2026
Same author

Diagnostic comparisons of PTSD and complex PTSD in clinical adolescents: Detection rates and ACEs as risk factors.

European journal of psychotraumatology·2026
Same author

Research on the decision-making of high-tech manufacturing enterprises for energy conservation and emission reduction driven by industrial green microgrids under the green credit regulatory system.

Scientific reports·2026
Same author

Smart double-screening system of propagating male-sterile lines for maize hybrid seed production.

Journal of integrative plant biology·2026
Same author

Glycogen Synthase Kinase-3β Inhibition Ameliorates Synaptic and Mitochondrial Dysfunction in a Sporadic Alzheimer's Disease-Like Model.

Molecular neurobiology·2026
Same author

Novel perspectives on PPARγ regulation: from SPPARMs to the emerging role of lncRNAs in metabolic disorders.

International journal of obesity (2005)·2026

Related Experiment Video

Updated: Sep 10, 2025

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

34.4K

An engineered CRISPR-Cas12i tool for efficient multiplexed genome editing.

Linli Wang1,2,3, Yanlu Wang1,2,3, Jian Chen4

  • 1State Key Laboratory of Animal Biotech Breeding, China Agricultural University, Beijing 100193, China.

Nucleic Acids Research
|August 28, 2025
PubMed
Summary

Researchers engineered Cas12i.3, a CRISPR RNA (crRNA)-guided nuclease, to improve multiplexed genome editing. The enhanced optimized Cas12i (EOCas12i) systems enable efficient editing of multiple targets simultaneously, offering a simplified tool for genetic research.

More Related Videos

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
10:18

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art

Published on: May 28, 2019

17.2K
Genome Editing in Mammalian Cell Lines using CRISPR-Cas
07:56

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

22.0K

Related Experiment Videos

Last Updated: Sep 10, 2025

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

34.4K
CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
10:18

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art

Published on: May 28, 2019

17.2K
Genome Editing in Mammalian Cell Lines using CRISPR-Cas
07:56

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

22.0K

Area of Science:

  • Molecular Biology
  • Genome Engineering
  • Biotechnology

Background:

  • Multiplexed genome editing tools face challenges in precursor CRISPR RNA (pre-crRNA) processing and require additional regulatory components.
  • The Cas12i.3 nuclease lacks pre-crRNA processing ability, limiting its multiplexing potential.

Purpose of the Study:

  • To engineer a highly efficient and simplified multiplexed genome editing system based on Cas12i.3.
  • To overcome the limitations of wild-type Cas12i.3 for broad genome editing applications.

Main Methods:

  • Optimized CRISPR RNA (crRNA) design, codon usage, and exonuclease fusion to create initial optimized Cas12i (IOCas12i).
  • Employed rational design and amino acid mutations to develop enhanced optimized Cas12i (EOCas12i) systems (EOCas12i-Combo1 and EOCas12i-Combo2).
  • Evaluated editing efficiency, specificity, and multiplexing capability using up to 30 target crRNA arrays.

Main Results:

  • Engineered EOCas12i systems demonstrated significantly enhanced editing efficiencies (2.5- to 60.0-fold) compared to wild-type Cas12i.3.
  • Achieved editing efficiencies comparable to established systems like Streptococcus pyogenes Cas9 (SpCas9) and Lachnospiraceae bacterium Cas12a (LbCas12a).
  • Demonstrated efficient multiplexed editing of up to 30 targets using compact crRNA arrays, producing longer indels beneficial for gene knockout.

Conclusions:

  • The developed EOCas12i-Combo1 and EOCas12i-Combo2 systems represent significant advancements in multiplexed genome editing.
  • These engineered nucleases offer a simplified and highly efficient platform for diverse genome editing applications.
  • The enhanced Cas12i variants overcome previous limitations, paving the way for broader use in genetic research and engineering.