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CRISPR

CRISPR

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...

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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.
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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...

The Antiviral System of Bacteria and Archaea: CRISPR

The 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...

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Updated: Jul 19, 2025

Substrate Generation for Endonucleases of CRISPR/Cas Systems

Substrate Generation for Endonucleases of CRISPR/Cas Systems

Published on: September 8, 2012

Kinetic dissection of pre-crRNA binding and processing by CRISPR-Cas12a.

Selma Sinan¹, Nathan M Appleby¹, Rick Russell¹

¹Department of Molecular Biosciences, University of Texas at Austin, Austin TX 78712.

Biorxiv : the Preprint Server for Biology

|August 7, 2023

Summary

CRISPR-Cas12a exhibits extremely tight binding to precursor CRISPR RNA (pre-crRNA), making binding the rate-limiting step for genome editing. This tight binding dictates Cas12a specificity and can be optimized through crRNA design.

Keywords:

AsCas12a CRISPR-Cas system RNA folding RNA-guided nuclease rate-limiting binding

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Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

CRISPR-Cas12a is a key enzyme in genome editing.
Cas12a processes single precursor CRISPR RNAs (pre-crRNAs) for its function.
Understanding the kinetics and thermodynamics of pre-crRNA processing is crucial for optimizing genome editing tools.

Conclusions:

Cas12a pre-crRNA binding kinetics and thermodynamics are quantitatively defined.
Pre-crRNA binding affinity and processing specificity are primarily determined by the guide sequence.
Strategies for optimizing crRNA design, including 5'-phosphorylation, can enhance Cas12a-mediated genome editing efficiency.