LbCas12a, His-Tagged

Alternate Names:

Cpf1 (synthetic construct)

UniProt Number:

A0A5S8WF58

Overview:

CRISPR (clustered regularly interspaced short palindromic repeats) and their CRISPR-associated (Cas) proteins constitute the adaptive immune system in bacteria (1-2). This system has been redesigned to create an exemplary genome editing tool for application to RNA-based therapeutics development (3) and disease diagnosis of viral pathogens. Cas12 and Cas13 are recognized as ideal candidates for future development and management of diseases such as cancer (4). Several CRISPR-Cas12-based SARS-CoV-2 detection assays have been developed (5). CRISPR/Cas12 (LbCpf1) is a Cas12 synthetic construct derived from Lachnospiraceae bacteria.

Accession Number:

ATB19154.1

Formulation:

Protein stored in 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, 150 mM imidazole, 1 mM DTT and 10% glycerol

References:

1. Horvath, P., et al. CRISPR/Cas, the immune system of bacteria and archaea. Science, 327(5962):167-170.
2. Morange, M. What history tells us XXXVII. CRISPR-Cas: The discovery of an immune system in prokaryotes. J. Biosci. 40(2):221-223.
3. Gier, R.A., et al. 2020. High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening. Nat. Commun. 11(1):1-9.
4. Singh, M., et al. 2022. The era of Cas12 and Cas13 CRISPR-based disease diagnosis. Critical Reviews in Microbiology, DOI: 10.1080/1040841X.2021.2025041.
5. Broughton JP, et al. 2020. Rapid detection of 2019 novel coronavirus SARS-CoV-2 using a CRISPR-based DETECTR lateral flow assay. medRxiv doi: https://doi.org/10.1101/ 2020.03.06.20032334.

Storage and Stability:

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

Concentration:

0.1 μg/μl
7.14 pmol/μg

Molecular Weight:

Calculated: 144.8 kDa
Observed: ~130 kDa

Purity:

>90%

Recommendation:

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