Description

Adaptive immune systems from prokaryotes utilize clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR associated (Cas) proteins to cleave foreign genetic material. Cas13a, (previously referred to as C2c2) is part of the Type VI CRISPR-Cas system. Cas13a functions similarly to Cas9, using a ~64-nt guide RNA to encode target specificity. The Cas13a protein complexes with the guide RNA via recognition of a short hairpin in the crRNA, and target specificity is encoded by a 28 – 30-nt spacer that is complementary to the target region. Cas13a cuts within the crRNA direct repeat in a pre-crRNA array to produce a mature Cas13a-crRNA complex.

In addition to programmable RNase activity, all Cas13s exhibit collateral activity after recognition and cleavage of a target transcript, leading to non-specific degradation of any nearby transcripts regardless of complementarity to the spacer. The RNA-activated RNA cleavage activity of Cas13a allows for RNA detection and potential diagnostic applications.

Applications • Targeted RNA knockdown • RNA editing • Nucleic acid detection

Source E. coli

Organism Leptotrichia wadeii (strain F0279)

UniProt # U2PSH1

Molecule Weight The mature recombinant Cas13a is a 143.7 kDa protein containing 1182aa.

Purity > 95 % as determined by SDS-PAGE

Storage -20 Avoid repeated freeze-thaw cycles.

Cas13 Storage Buffer 0.3 M NaCl 20 mM Tris-HCl, pH 8.0 0.5 mM EDTA 1 mM DTT 50% Glycerol

10X Cas13 Reaction Buffer 200 mM HEPES, 50 mM MgCl2, 1 M NaCl, 1 mM EDTA, pH 6.5

10X Cas13 sherlock reaction buffer (requested) 200 mM HEPES, 90 mM MgCl2, pH 6.8

Citations & Technical Literature

1. Nature, Vol. 613, pages 588 – 594 (2023) / Read Article Cas12a2 elicits abortive infection through RNA-triggered destruction of dsDNA Oleg Dmytrenko, et al. “LwCas13a was purchased from MCLAB Molecular Cloning Laboratories (Cas13a-100). Reactions (10 μl) containing 250 nM of LwCas13a and 500 nM of its cognate crRNA in the provided 1× Cas9 buffer”

2. Microbiology Spectrum, Vol. 10, No. 5 (2022)/ Read Article Reverse Transcription-Loop-Mediated Isothermal Amplification CRISPR-Cas13a Technology as a Promising Diagnostic Tool for SARS-CoV-2 Concha Ortiz-Cartagena, et al. “0.15 μL Cas13a endonuclease (25 nM; MCLAB), 0.5 μL crRNA (50 nM; IDT), 2 μL reporter (1,000 nM; IDT), and 5 μL of a cDNA sample, adjusted to a final volume of 20 μL with dH2O”

3. Frontiers in Cellular and Infection Microbiology, Vol. 11 (2021)/ Read Article A Saliva-Based RNA Extraction-Free Workflow Integrated With Cas13a for SARS-CoV-2 Detection Iqbal Azmi, et al. “SHERLOCK assay of the above reaction mix was performed in a separate 1.5 ml Eppendorf tube which contains 1 μL of the Cas13a either from extracted pool or commercially (MCLAB, USA, Cat no. CAS13a-200) obtained source (at 63.3 μg/ml concentration),”

COA ( Certificate of Analysis )