|
Description
CRISPR-Cas systems are powerful molecular tools for mediating nucleic acids. Cas12a2, a Type V CRISPR/Cas nuclease, cleaves the target RNA following target RNA recognition by the complementary crRNA. Cas12a2 processes its own crRNAs. The PFS requirement for Cas12a2 is 5′-GAAAG-3′. Cas12a2 is able to indiscriminately degrade single-stranded DNA (ssDNA), double -stranded DNA (dsDNA) and single -stranded RNA (ssRNA) as collateral activities once sensing correct crRNA-guided RNA binding. These properties enable Cas12a2 as a single-effector nuclease efficiently and sensitively for RNA detection.
SuCas12a2 from Sulfuricurvum sp. PC08-66 is a programmable nuclease guided by a 42-44 nt crRNA and can be applied in RNA detection and genome editing.
Features
Low Endotoxin Level
High Biological Activity
Application
RNA Detection, CRISPR Diagnostics, Genome Editing
Source
Cas12a2 gene from Sulfuricurvum sp. PC08-66 (SuCas12a2, UniProtKB: A0A0C2W1L1) expressed in E. coli
Purity
Greater than 95% as determined by SDS-PAGE and FPLC
Activity
For the cleavage assay: 20 µl reaction, including SuCas12a2 Enzyme, crRNA, target RNA and 1x reaction buffer, preheated at 37°C for 15 min. Then for the ssDNA probe cleavage assay as Fig.1 below, added 100 nM ssDNA 5’FAM probe, incubated at 37°C for 30 min; For the plasmid cleavage assay as Fig. 2 below, added 7 nM pUC19 plasmid, incubated at 37°C for 25 min, and stop reaction with 1 µl Proteinase K (800 U/ml).
Storage Condition
-20 °C
10 X Reaction Buffer
500 mM Tris-HCl, pH 7.9
1M NaCl
100 mM MgCl2
1 mg/ml BSA
Storage Buffer
25 mM HEPES, pH 7.2
150 mM NaCl
2 mM MgCl2
50% glycerol
Experimental Data
References
1. Oleg Dmytrenko, et. al. (2023). Nature. 613, 588 -– 594
2. Jack P. K. Bravo, et. al. (2023). Nature. 613, 582 -– 587
|
