Related Experiment Video
Updated: Dec 21, 2025
11:53
Substrate Generation for Endonucleases of CRISPR/Cas Systems
Published on: September 8, 2012
27.9K
Purification of specific DNA species using the CRISPR system
Toshitsugu Fujita1, Hodaka Fujii1
1Department of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, Zaifu-cho 5, Hirosaki, Aomori, Japan.
Biology Methods & Protocols
|May 13, 2020
Summary
Researchers developed CRISPR-mediated DNA purification for identifying associated molecules. This technology enables precise isolation of specific DNA sequences within chromatin structures for advanced analysis.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Chromatin immunoprecipitation (ChIP) is a standard technique for studying DNA-protein interactions.
- Limitations exist in ChIP's specificity and efficiency for isolating target DNA sequences within complex chromatin structures.
Purpose of the Study:
- To introduce and review the principles and advancements of CRISPR-mediated DNA purification.
- To highlight the utility of this method for identifying DNA-associated molecules and enriching specific DNA sequences.
Main Methods:
- Development of engineered DNA-binding molecules incorporating the clustered regularly interspaced short palindromic repeats (CRISPR) system.
- Application of CRISPR complexes for in-cell or in vitro tagging of target DNA sequences.
- Utilizing CRISPR ribonucleoprotein complexes with nuclease-dead Cas9 (dCas9) and synthetic guide RNA for in vitro tagging.
Main Results:
- Successful purification of specific DNA species within chromatin structures.
- Enabling identification of associated proteins, RNAs, and other genomic regions.
- Demonstration of enrichment of purified DNA sequences for applications like next-generation sequencing.
Conclusions:
- CRISPR-mediated purification offers a powerful and versatile approach for targeted DNA isolation.
- This technology enhances the study of DNA-associated molecules and facilitates genomic enrichment strategies.
- Ongoing advancements continue to expand the applications of CRISPR-based DNA purification.
Related Concept Videos
01:28CRISPR/Cas9 Genome Editing
1.4K
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...
1.4K
01:59CRISPR
57.2K
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...
57.2K
02:53CRISPR and crRNAs
18.5K
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...
18.5K
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
02:31Homologous Recombination
61.9K
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...
61.9K
01:23The Antiviral System of Bacteria and Archaea: CRISPR
523
CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
523