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Theclustered regularly interspaced short palindromic repeats, known as CRISPR systems are adaptive immune mechanisms commonlypresent in archaea and bacteria. The CRISPR systems enable the host to specificallytarget and cleave foreign nucleic acids thus targeting infectiousviruses andplasmids. Recently, a type V CRISPR system has been identified in severalbacteria, the Cpf1 CRISPR from Prevotella and Francisella 1. In contrast toCas9 systems, CRISPR/Cpf1 systems are smaller in size, do not require anadditional trans-activating crRNA (tracrRNA), and allow for targeting ofadditional genomic regions by cleaveing the target DNA proceeded by a shortT-rich protospacer-adjacent motif (PAM). On the other hand, the Cas9 system requiresa G-rich PAM following the target DNA. Furthermore, Cas12a/Cpf1 introduces astaggered DNA double stranded break with a 4 or 5-nt 5’ overhang. RecombinantAcidaminococcus sp. BV3L6 Cas12a (cpf1) nuclease is expressed in E. coli andpurified. The nuclease contains nuclear localization sequence (NLS) at theC-terminus and 6× His-tag at the C-terminus.
Note
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.
Applications
crRNA -dependentdouble-stranded DNA cleavage
Properties
Expression System
RecombinantCas12a with an C-terminal NLS expressed by E.coli
Species
Acidaminococcus sp. (strain BV3L6)
Tag
C-terminal 6× His Tag
Apparent Molecular Weight
~150kDa, on SDS-PAGE under reducing conditions
Concentration
4 mg/ml
Active temperature
ThisCas12a is active at 37℃.
Formulation
Suppliedas a solution of 10 mM Tris, pH7.4,300 mM NaCl, 0.5 mM DTT, and 50%glycerol.
Storage & Stability
This product remains stable up to 18 months at -20℃. Avoid repeated freeze-thaw cycles.
Accession#
U2UMQ6
Key Features
High knockoutefficiencies: Consistent high performance in in-vitro plasmid cleavage test.Time-saving:noneed for transcription and translation.DNA-free:noexternal DNA added to system.