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GenCRISPR™ Cas12a (Cpf1) Nuclease
GenCRISPR™ Cas12a (Cpf1) Nuclease
Key functions:
- DNA editing ss or ds
- PAM: 5′-TTTV-3′ upstream
- AT rich region editing
The
clustered regularly interspaced short palindromic repeats, known as CRISPR systems are adaptive immune mechanisms commonly
present in archaea and bacteria. The CRISPR systems enable the host to specifically
target and cleave foreign nucleic acids thus targeting infectiousviruses and
plasmids. Recently, a type V CRISPR system has been identified in several
bacteria, the Cpf1 CRISPR from Prevotella and Francisella 1. In contrast to
Cas9 systems, CRISPR/Cpf1 systems are smaller in size, do not require an
additional trans-activating crRNA (tracrRNA), and allow for targeting of
additional genomic regions by cleaveing the target DNA proceeded by a short
T-rich protospacer-adjacent motif (PAM). On the other hand, the Cas9 system requires
a G-rich PAM following the target DNA. Furthermore, Cas12a/Cpf1 introduces a
staggered DNA double stranded break with a 4 or 5-nt 5’ overhang. Recombinant
Acidaminococcus sp. BV3L6 Cas12a (cpf1) nuclease is expressed in E. coli and
purified. The nuclease contains nuclear localization sequence (NLS) at the
C-terminus and 6× His-tag at the C-terminus. |
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.
| Expression System |
Recombinant
Cas12a with an C-terminal NLS expressed by E.coli |
| Species |
Acidaminococcus sp. (strain BV3L6) |
| Tag |
C-terminal 6× His Tag |
| Apparent Molecular Weight |
~150
kDa, on SDS-PAGE under reducing conditions |
| Concentration |
4 mg/ml |
| Active temperature |
This
Cas12a is active at 37℃. |
| Formulation |
Supplied
as a solution of 10 mM Tris, pH7.4,
300 mM NaCl, 0.5 mM DTT, and 50%
glycerol. |
| Storage | This product remains stable up to 18 months at -20℃. Avoid repeated freeze-thaw cycles. |
| Accession# |
U2UMQ6 |
| Key Features |
High knockout
efficiencies: Consistent high performance in in-vitro plasmid cleavage test. Time-saving: no need for transcription and translation. DNA-free: no external DNA added to system. |
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| Quality Control Specifications |
|
|
crRNA -dependent
double-stranded DNA cleavage |
A 20 μl reaction in 1xCas12a Nuclease Reaction Buffer containing 60 ng linearized plasmid, 10 ng crRNA, and 100 ng GenCRISPR™ Cas12a (Cpf1) Nuclease for 30 mins at 37°C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.
For more documents, please visit "Technical Support".
