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