TccCas13a

Alternate Names:

Type VI-A CRISPR-associated RNA-guided ribonuclease Cas13a

Overview:

CRISPR-Cas systems constitute the adaptive immune system in bacteria via RNA-guided endonucleases (1). Cas13a is an RNA-guided RNA endonuclease that belongs to the Class 2 type VI CRISPR-Cas system (2). Cas13a possess both RNA recognition and cleavage activities and cleaves RNA through conserved basic residues within its two HEPN domains (3). Thermoclostridium caenicola Cas13a is thermostable and exhibits robust cis and trans catalytic activities over a broad range of temperatures from 37 to 70 °C (4). This enzyme may be used for various biotechnological applications including diagnostics development, transcriptome engineering, editing, and targeted therapeutics.

Accession Number:

WP_149678719

Formulation:

Protein stored in 50 mM sodium phosphate, pH 7.5, 150 mM imidazole, 1 mM DTT, 10% glycerol

References:

1. Horvath, P., et al. 2010. CRISPR/Cas, the immune system of bacteria and archaea. Science, 327(5962):167-170.
2. East-Seletsky A, et al. 2016. Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 538:270–3.
3. Abudayyeh O, et al. 2016. C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 353:5573.
4. Mahas A, et al. 2022. Characterization of a thermostable Cas13 enzyme for one-pot detection of SARS-CoV-2. Proc Natl Acad Sci U S A. 119(28):e2118260119.

Storage and Stability:

Store at –70°C. To avoid repeated handling and multiple freeze/thaw cycles, aliquot product into smaller quantities.

Concentration:

0.2 μg/μl
6.91 pmol/μg

Purity:

>95%

Molecular Weight:

Calculated: 144.7 kDa
Observed: ~139 kDa

Recommendation:

Always include RNase inhibitor at recommended concentrations in Cas enzyme reactions.