Related Experiment Video
Updated: Aug 5, 2026
10:46
Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
Published on: October 18, 2022
DIRECTOR: DNA-guided spatially ordered assembly of split crRNA for activating CRISPR/Cas12a
Lulu Qin1, Zhicheng Tang1, Yuehong Yu2
1School of Pharmacy, Nanjing Medical University, Nanjing, Jiangsu, 211166, China.
Biosensors & Bioelectronics
|August 3, 2026
Summary
A new DNA-guided strategy called DIRECTOR enhances CRISPR/Cas12a (CRISPR-associated protein 12a) activity for molecular diagnostics. It precisely regulates Cas12a activation, improving detection limits for nucleic acids.
Area of Science:
- Biotechnology
- Molecular Biology
- CRISPR Technology
Background:
- Precise regulation of Cas12a is vital for molecular diagnostics.
- Existing split CRISPR RNA (crRNA) systems lack activation efficiency and regulated mechanisms, hindering practical applications.
Purpose of the Study:
- To develop a novel DNA-guided strategy for activating CRISPR/Cas12a (CRISPR-associated protein 12a).
- To elucidate the regulatory mechanism of split crRNA assembly for Cas12a activation.
Main Methods:
- Utilized artificial intelligence (AlphaFold3) for structure prediction.
- Employed molecular dynamics simulations and fluorescence analysis.
- Investigated DNA-guided spatially ordered assembly of split crRNA.
Main Results:
- The 3' terminal extension of activator acts as a spatial director, stabilizing Cas12a domains and enabling activation.
- The 5' terminal extension acts as a spatial misdirector, inhibiting activation.
- Identified structural and energy thresholds for effective Cas12a activation.
- Achieved a limit of detection as low as 42.1 fM for miR-155 and dual-response detection.
Conclusions:
- The DIRECTOR strategy offers direct activation of Cas12a via DNA-guided assembly.
- Provides mechanistic insights for developing affordable and programmable CRISPR-based molecular diagnostics.
Related Concept Videos
01:59CRISPR
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 Short...
01:59CRISPR
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 Short...
01:28CRISPR/Cas9 Genome Editing
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...
02:53CRISPR and crRNAs
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...
01:23The Antiviral System of Bacteria and Archaea: CRISPR
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 defense.
02:31Homologous Recombination
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...