Related Experiment Video
Updated: Dec 6, 2025
07:22
Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans
Published on: May 23, 2020
9.6K
An efficient KRAB domain for CRISPRi applications in human cells
Nader Alerasool1,2, Dmitri Segal1,2, Hunsang Lee1
1Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada.
Nature Methods
|October 6, 2020
Summary
Clustered regularly interspaced short palindromic repeat interference (CRISPRi) offers gene silencing but can be incomplete. Researchers identified the ZIM3 KRAB domain, significantly enhancing CRISPRi
Area of Science:
- Molecular Biology
- Gene Regulation
- Biotechnology
Background:
- Clustered regularly interspaced short palindromic repeat interference (CRISPRi) utilizes dCas9-KRAB for gene silencing.
- Existing CRISPRi systems often exhibit incomplete repression of target genes.
Purpose of the Study:
- To identify and characterize highly potent KRAB domains for improved CRISPRi gene silencing.
- To develop a more efficient gene silencing platform using enhanced CRISPRi.
Main Methods:
- Assayed 57 different Krüppel-associated box (KRAB) domains for their repressive potency.
- Constructed and tested dCas9 fused to the most potent KRAB domains.
Main Results:
- Identified the ZIM3 KRAB domain as a significantly more potent repressor compared to others.
- Demonstrated that ZIM3 KRAB-dCas9 fusion achieves superior gene silencing efficiency.
Conclusions:
- The ZIM3 KRAB domain represents a substantial improvement for CRISPRi-mediated gene silencing.
- This enhanced CRISPRi platform offers greater precision and effectiveness in controlling gene expression.
Related Concept Videos
02:53CRISPR and crRNAs
18.3K
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...
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...
18.3K
01:59CRISPR
56.1K
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...
56.1K
02:53Conservative Site-specific Recombination and Phase Variation
6.5K
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...
The recognition sites for Cre recombinase called LoxP...
6.5K