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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...

CRISPR and crRNAs

CRISPR 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...

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Updated: Jun 13, 2025

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Metagenome-Derived CRISPR-Cas12a Mining and Characterization.

Kalani Gast¹, Sydney Baker¹, Adair L Borges²

¹Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, North Carolina, USA.

The CRISPR Journal

|May 21, 2025

Summary

Researchers discovered novel CRISPR-Cas12a enzymes in dairy cow microbes, enhancing genome editing tools. Mutagenesis further improved the efficiency of these Cas12a variants for gene editing applications.

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Published on: February 26, 2019

Area of Science:

Genetics and Genomics
Molecular Biology
Microbiology

Background:

CRISPR-Cas systems, particularly CRISPR-Cas12a, are powerful tools for genome editing.
There is ongoing research to expand the CRISPR-Cas toolbox with novel and efficient effectors.
Metagenomic data offers a rich source for discovering new CRISPR-associated proteins (Cas).

Purpose of the Study:

To discover and characterize novel CRISPR-Cas12a enzymes from dairy cow microbial metagenomes.
To assess the genome editing potential of newly identified Cas12a systems.
To investigate sequence variations for improving Cas12a activity.

Main Methods:

Bioinformatic analysis of dairy cow microbial metagenomes to identify CRISPR-Cas systems.
In silico characterization of guide RNAs and protospacer adjacent motifs for Cas12a systems.
In vitro assessment using cell-free transcription-translation assays with GFP readouts.
In vivo genome editing evaluation in Escherichia coli using 1 kb knockouts.

Main Results:

Identification and characterization of five novel CRISPR-Cas12a systems.
Demonstration of genome editing capabilities, including 1 kb knockouts in E. coli.
Discovery of natural sequence variation in the bridge-helix domain of a top-performing Cas12a.
Mutagenesis of Cas12a orthologs led to enhanced gene editing efficiency in an underperforming candidate.

Conclusions:

Dairy cow metagenomes harbor undiscovered CRISPR-Cas12a enzymes with genome editing potential.
Sequence variation, particularly in the bridge-helix domain, can be leveraged to refine Cas12a activity.
Exploration of metagenomic diversity is a promising strategy for developing advanced genome editing tools.