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EnGen® Lba Cas12a (Cpf1)

Catalog # Concentration Size List Price Quantity Your Price
M0653S 1 µM 70 pmol $107.00
$96.30
M0653T 100 µM 2000 pmol $381.00
$342.90
Catalog # Size List Price Your Price
M0653S 70 pmol $107.00
$96.30
M0653T 2000 pmol $381.00
$342.90
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

EnGen Lba Cas12a (Cpf1) is a programmable DNA endonuclease guided by a single guide RNA (gRNA). Targeting requires a gRNA complementary to the target site as well as a 5' TTTV protospacer adjacent motif (PAM) on the DNA strand opposite the target sequence. Cleavage by EnGen Lba Cas12a (Cpf1) occurs ~17 bases 3' of the PAM and leaves 5' overhanging ends.

  • T-rich TTTV PAM sequence opens up additional genomic regions for targeting
  • Shorter, 40-44 base guide RNA
  • Two nuclear localization signals for improved transport to the nucleus
  • 5’ overhanging termini on cleavage products
  • Active from 16 to 48°C
  • Maintains activity at lower temperatures than the Acidaminococcus orthologs, permitting editing in ectothermic organisms such as zebra fish and xenopus
  • High concentration liquid format can be used for microinjection, electroporation and lipofection.

EnGen® Lba Cas12a from Lachnospiraceae bacterium ND2006 is a site-specific DNA endonuclease guided by a single 41-44 nucleotide guide RNA (gRNA) (1). Targeting requires a gRNA complementary to the target site as well as a 5’ TTTV protospacer adjacent motif (PAM) on the DNA strand opposite the target sequence. Cleavage by EnGen Lba Cas12a occurs ~18 bases 3′ of the PAM and leaves 5′ overhanging ends. EnGen Lba Cas12a has Simian virus 40 (SV40) T antigen nuclear localization sequences (NLS) at both the N and C-termini of the protein.

 

Schematic representation of Lba Cas12a nuclease sequence recognition and DNA cleavage

Cas12a


Expanded temperature range of Lba Cas12a

Cas12a Temperature

Purified Lba or Asp Cas12a proteins were used to form RNP complexes with their canonical guide RNAs. The DNA target is a single a PCR product 5´-end labeled with FAM. Lba or Asp Cas12a RNPs were incubated with target DNA at the temperatures indicated at a ratio of 10:1 RNP to DNA for 10 minutes. Cleavage products were resolved by capillary electrophoresis and quantitated by integration of signal corresponding to intact and cleaved fragments. Data are shown as the mean +/- SD of duplicate experiments.
Product Source
EnGen Lba Cas12a from Lachnospiraceae bacterium ND2006 is expressed as a N-terminal 6XHis-tagged fusion in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  EnGen® Lba Cas12a (Cpf1) M0653SVIAL -20 1 x 0.07 ml 1 µM
  NEBuffer™ r2.1 B6002SVIAL -20 1 x 1.25 ml 10 X
  EnGen® Lba Cas12a (Cpf1) M0653TVIAL -20 1 x 0.02 ml 100 µM
  EnGen Lba Cas12a Diluent B0653AVIAL -20 2 x 1 ml 1 X
  NEBuffer™ r2.1 B6002SVIAL -20 1 x 1.25 ml 10 X

Properties & Usage

Reaction Conditions

1X NEBuffer™ r2.1
Incubate at 37°C

1X NEBuffer™ r2.1
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Storage Buffer

500 mM NaCl
20 mM sodium acetate
0.1 mM EDTA
0.1 mM TCEP
50% Glycerol
pH 6 @ 25°C

Heat Inactivation

65°C for 10 minutes

Notes
  • 100 µM is equal to 15.1 mg/ml EnGen Lba Cas12a.
References
  • Zetsche, B., et. al. (2015). Cell. 163, 759-771.
  • Moreno-Mateos, M.A., et al. (2017). Nat. Commun.. 8, 2024. PubMedID: 29222508
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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