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Description
CRISPR-Cas systems are powerful tools for mediating nucleic acids. Cas12a (also referred as Cpf1) nuclease, belonging to the Class 2, Type V CRISPR system, is a compact and efficient enzyme that cleaves the target double-stranded DNA into staggering ends. Cas12a processes its own mature crRNA. The PAM requirement for Cas12a is 5′-TTTN. Cas12a is able to indiscriminately cleave single-stranded DNA once sensing correct RNA-guided DNA binding. This property enables Cas12a as an efficient, rapid and sensitive DNA Detection tool besides engineered as a platform for Genome Editing.
LbCas12a from Lachnospiraceae bacterium (strain ND2006) is a programmable DNA endonuclease guided by a 42-44nt crRNA and can be applied in vitro DNA digestion and DNA detection.
LbCas12a-NLS is fused with both N- and C-terminal SV40 (simian virus 40) large T antigen nuclear localization signals (NLSs) for improved transport to the nucleus to heterologous expressed in mammalian and plant cells for cleavage of either exogenous or endogenous nucleic acids.
Features
Low Endotoxin Level
High Biological Activity
Application
Genome Editing, DNA Detection, CRISPR Diagnostics
Source
Cas12a gene from Lachnospiraceae bacterium (strain ND2006) expressed in E. coli
Purity
Greater than 98% as determined by SDS-PAGE
Activity
97% of PCR product digested after 1 h reaction at 37 °C.
The reaction is performed as follow: 30µl reaction, including 1 µl Cas12a Enzyme (1µM), 100ng crRNA, 50 ng purified PCR product and 1x reaction buffer. Incubate 1 h at 37°C, and stop reaction with 1µl Proteinase K (800 U/ml).
Storage condition
-20 °C
10 X Reaction buffer
500 mM NaCl
100 mM Tris-HCl
100 mM MgCl2
1 mg/ml Bovine Serum Albumin (BSA) Ultrapure
(pH 7.9 at 25 °C)
Storage Buffer
500 mM NaCl
20 mM sodium acetate
0.1 mM EDTA
0.1 mM TCEP
50% Glycerol
(pH 6 at 25 °C)
Experimental Data
References
1. Zetsche, B., et. al. (2015). Cell. 163, 759-771.
2. Chen, J.S., et. al. (2018). Science. 360, 436-439.
3. Abudayyeh, O.O. and Gootenberg, J.S. (2021). Science. 372 (6545), 914-915.
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