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

72
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...
72
CRISPR01:59

CRISPR

52.4K
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...
52.4K
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
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
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...
6.0K

You might also read

Related Articles

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

Sort by
Same author

Convergent mechanisms in Wnt and Hedgehog signaling.

Science signaling·2026
Same author

Combinatorial and Inducible CRISPRa/i Enables Canalized hiPSC Forward Programming and Iterative Refinement <i>via</i> Single-Cell Genomics.

bioRxiv : the preprint server for biology·2026
Same author

Auxin promotes robust founder cell specification during Arabidopsis lateral root initiation.

Genetics·2026
Same author

Bioproduction, bioprotection, and biocontainment in multi-kingdom microbial systems with 3D spatial control.

Biofabrication·2026
Same author

The response to kidney injury is epigenetically regulated through the activation of bivalent genes.

American journal of physiology. Renal physiology·2026
Same author

Cell-Free-Based Thermophilic Biocatalyst for the Synthesis of Amino Acids from One-Carbon Feedstocks.

ACS synthetic biology·2025

Related Experiment Video

Updated: Jul 23, 2025

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
10:46

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

1.8K

Engineering activatable promoters for scalable and multi-input CRISPRa/i circuits.

Diego Alba Burbano1,2, Ryan A L Cardiff2,3, Benjamin I Tickman2,3

  • 1Department of Chemical Engineering, University of Washington, Seattle, WA 98195.

Proceedings of the National Academy of Sciences of the United States of America
|July 18, 2023
PubMed
Summary

Scientists engineered high-performance activatable promoters for building complex gene regulatory networks (GRNs) using CRISPR technology. This breakthrough enables deeper and wider synthetic GRNs with precise control in cell-free systems.

Keywords:
CRISPR activationactivatable promoter engineeringcell-freegene regulatory networksgenetic circuits

More Related Videos

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
10:46

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

Published on: June 2, 2018

9.4K
A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
08:20

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

4.2K

Related Experiment Videos

Last Updated: Jul 23, 2025

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
10:46

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

1.8K
Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
10:46

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines

Published on: June 2, 2018

9.4K
A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
08:20

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

4.2K

Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Biotechnology

Background:

  • Gene regulatory networks (GRNs) are fundamental to biological systems.
  • CRISPR-based genetic circuits offer a powerful platform for constructing synthetic GRNs.
  • Developing high-performance components is crucial for advanced GRN engineering.

Purpose of the Study:

  • To develop an approach for creating high-performing activatable promoters for CRISPR-activation and -interference (CRISPRa/i) gene regulatory networks.
  • To enable the construction of deep, wide, and multi-input synthetic GRNs.
  • To demonstrate the generalizability and application of engineered promoters in cell-free systems.

Main Methods:

  • Integrated sequence-based design and in vivo screening to engineer activatable promoters.
  • Utilized an *Escherichia coli*-based cell-free system for promoter characterization.
  • Assembled engineered promoters into multilayer CRISPRa/i GRNs.

Main Results:

  • Achieved up to 1,000-fold dynamic range in engineered activatable promoters.
  • Demonstrated the construction of CRISPRa GRNs six layers deep and four branches wide.
  • Improved the dynamic range of the EL222 optogenetic system from 6-fold to 34-fold.
  • Showcased applications in small molecule and protein-protein interaction-mediated CRISPRa systems.

Conclusions:

  • Developed a generalizable method for designing high dynamic range activatable promoters.
  • Enabled novel gene regulatory functions, including feedback loops and logic gates, in cell-free systems.
  • Advanced the potential of CRISPR-based synthetic biology for complex biological circuit construction.