Using Cpf1 (a.k.a. Cas12a) in your CRISPR experiment offers several advantages over other CRISPR-associated nucleases.
Due to the T-rich PAM sequence (TTTN), Cpf1 enables editing in regions unable to be targeted by Cas9.
Cpf1 can be used with a shorter guide RNA (called crRNA) than Cas9.
Cpf1 creates a staggered cut in dsDNA instead of a blunt cut.
Cpf1 cuts distal to the PAM sequence, which may allow for multiple rounds of cleavage.
asCpf1 is from the bacteria
Acidaminococcus
. This protein contains a SV40 T antigen nuclear localization signal (NLS) on the N-terminus of the protein. If the cut caused by asCpf1 is repaired by non-homologous end joining (NHEJ), an indel may be formed that disrupts the open reading frame of the targeted gene, leading to gene knockout. Alternatively, by supplying a repair template, a sequence can be knocked in at the cleavage site via homology directed repair (HDR).
SKU
K088
Cas Type
Nuclease
Cas Origin
asCpf1
Cas Protein Marker
No GFP
Source
E. coli
Alias
Cas12a, asCas12a, Cpf1 from Acidaminococcus, Cas12a from Acidaminococcus
Unit quantity
38 μg (250 pmol/ 25µL)
Product Concentration
10 μM, 1.5 mg/ml
Format
Enzyme supplied with 10X Reaction Buffer.
Storage Buffer
10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 300 mM NaCl, and 50% (v/v) Glycerol.
Storage
Store all components at -20°C.
Caution
This product is distributed for laboratory research only.
Caution: Not for diagnostic use.
Nelson, C. E., Wu, Y., Gemberling, M. P., Oliver, M. L., Waller, M. A., Bohning, J. D., … Gersbach, C. A. "Long-term evaluation of AAV-CRISPR genome editing for Duchenne muscular dystrophy" Nature Medicine 25(3):427–432 (2019). DOI: 10.1038/s41591-019-0344-3.
