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

3.2K
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...
3.2K
CRISPR01:59

CRISPR

46.7K
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...
46.7K
CRISPR01:59

CRISPR

13.9K
13.9K
What is Genetic Engineering?00:49

What is Genetic Engineering?

71.0K
Overview
71.0K
CRISPR and crRNAs02:53

CRISPR and crRNAs

14.7K
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...
14.7K
Homologous Recombination02:31

Homologous Recombination

59.0K
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...
59.0K

You might also read

Related Articles

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

Sort by
Same author

Placental growth factor (PLGF)-based testing to help diagnose suspected pre-eclampsia: a systematic review and economic evaluation.

Health technology assessment (Winchester, England)·2026
Same author

Effects of single-injection vs. continuous brachial plexus blocks for shoulder surgeries on patient-reported outcomes: a systematic review and meta-analysis with trial sequential analysis of randomised controlled trials.

Anaesthesia·2026
Same author

Postoperative Delirium and Neurocognitive Disorders Associated With Inflammatory and Neuronal Injury Biomarkers: A Prospective, Longitudinal, Cohort Study.

Anesthesia and analgesia·2026
Same author

Routine pre-operative modified telephone interview for cognitive status screening: a reply.

Anaesthesia·2026
Same author

Intraoperative electroencephalogram-derived measures and their association with postoperative delirium. Response to Br J Anaesth 2026; 136: 1055-56.

British journal of anaesthesia·2026
Same author

LncRNA-HLX-2-7/HLX axis-dependent metabolic reprogramming drives cancer progression in group 3 medulloblastoma.

Acta neuropathologica communications·2026
Same journal

An assay to quantify sexual commitment and stage conversion in the human malaria parasite Plasmodium falciparum.

Nature protocols·2026
Same journal

Author Correction: Direct inoculation of bioreactor-controlled stirred suspension culture with cryopreserved human pluripotent stem cells.

Nature protocols·2026
Same journal

High-throughput measurements of protein domain functions using magnetic separation.

Nature protocols·2026
Same journal

Inducing physiological polarity and performing gene editing using CRISPR-Cas9 in human trophoblast organoids.

Nature protocols·2026
Same journal

Photocatalytic low-temperature defluorination of PTFE.

Nature protocols·2026
Same journal

Multimodal imaging and quantification of lanthanide chelate-labeled micro- and nanoplastics in plants.

Nature protocols·2026
See all related articles

Related Experiment Video

Updated: May 6, 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

36.0K

Genome engineering using the CRISPR-Cas9 system.

F Ann Ran1,2,3,4,5, Patrick D Hsu1,2,3,4,5, Jason Wright1

  • 1Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, Massachusetts, USA.

Nature Protocols
|October 26, 2013
PubMed
Summary
This summary is machine-generated.

This study introduces CRISPR-Cas9 genome editing tools for precise DNA modification in mammalian cells. It details methods for gene editing via nonhomologous end joining or homology-directed repair, including a double-nicking strategy to reduce errors.

More Related Videos

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

21.6K
Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
09:04

Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells

Published on: September 25, 2019

7.4K

Related Experiment Videos

Last Updated: May 6, 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

36.0K
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

21.6K
Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
09:04

Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells

Published on: September 25, 2019

7.4K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Targeted nucleases enable precise genome alteration.
  • CRISPR-Cas9 system facilitates efficient genome engineering in eukaryotic cells using guide RNA specificity.

Purpose of the Study:

  • To describe tools for Cas9-mediated genome editing in mammalian cells via NHEJ and HDR.
  • To present methods for generating modified cell lines for functional studies.
  • To introduce a double-nicking strategy using Cas9 nickase and paired guide RNAs to minimize off-target cleavage.

Main Methods:

  • Utilized CRISPR-Cas9 system for genome editing.
  • Employed nonhomologous end joining (NHEJ) and homology-directed repair (HDR) pathways.
  • Implemented a double-nicking strategy with Cas9 nickase and paired guide RNAs for enhanced specificity.

Main Results:

  • Provided guidelines for target site selection, cleavage efficiency evaluation, and off-target activity analysis.
  • Achieved gene modifications within 1-2 weeks.
  • Derived modified clonal cell lines within 2-3 weeks.

Conclusions:

  • The described CRISPR-Cas9 tools and strategies enable efficient and precise genome editing in mammalian cells.
  • The double-nicking approach effectively minimizes off-target modifications.
  • This protocol accelerates the generation of genetically modified cell lines for downstream research.