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

97
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...
97
CRISPR01:59

CRISPR

52.6K
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...
52.6K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.1K
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...
6.1K
Homologous Recombination02:31

Homologous Recombination

50.8K
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...
50.8K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.1K
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...
17.1K

You might also read

Related Articles

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

Sort by
Same author

Mouse models to study von Willebrand factor in inflammation: a scoping review.

Intensive care medicine experimental·2026
Same author

Derivation of elephant induced pluripotent stem cells.

Nature methods·2026
Same author

Fully Modified SpyCas9 Guide RNAs Enable Robust Genome Editing In Cells and In Vivo.

bioRxiv : the preprint server for biology·2026
Same author

Structural basis of RNA-guided DNA integration by type I CRISPR-associated transposases.

bioRxiv : the preprint server for biology·2026
Same author

Transposon end recognition and excision mechanisms of type I-F CRISPR-associated transposases.

bioRxiv : the preprint server for biology·2026
Same author

Prime assembly with linear DNA donors enables large genomic insertions.

Nature·2026

Related Experiment Video

Updated: Aug 6, 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

34.2K

PAM-Flexible Genome Editing with an Engineered Chimeric Cas9.

Sabrina Koseki1, Lauren Hong1, Vivian Yudistyra1

  • 1Duke University.

Research Square
|March 22, 2023
PubMed
Summary

Researchers engineered a new CRISPR enzyme, SpRYc, by combining two Cas9 variants. This novel enzyme broadens genome editing capabilities by targeting diverse PAM sequences, enabling precise gene modification for therapeutic applications.

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

21.9K
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
11:35

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

Published on: June 16, 2017

12.7K

Related Experiment Videos

Last Updated: Aug 6, 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

34.2K
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

21.9K
Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
11:35

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

Published on: June 16, 2017

12.7K

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genomics

Background:

  • CRISPR-Cas9 systems require specific protospacer adjacent motifs (PAMs) for DNA targeting.
  • Limited PAM accessibility restricts the application of CRISPR genome editing.

Approach:

  • Engineered a chimeric Cas9 enzyme, SpRYc, by fusing the PAM-interacting domain of SpRY with the N-terminus of Sc++.
  • SpRY has an NRN > NYN PAM preference, while Sc++ offers broad, efficient, and accurate NNG editing.

Key Points:

  • SpRYc exhibits a highly flexible PAM preference, enabling editing of diverse NNN PAMs.
  • Demonstrated specific editing of disease-related loci using SpRYc.
  • The chimeric enzyme leverages the strengths of both parent Cas9 variants.

Conclusions:

  • Integrative protein design is a powerful strategy for advancing Cas9 engineering.
  • SpRYc's flexibility opens new avenues for precise genomic positioning in therapeutic applications.