Product Class: Other

EnGen® Lba Cas12a (Cpf1)
icon_recomb icon_neb2_1 icon_incTemp37 icon_HI_65
Item# Description List Price Web Price Qty
M0653S EnGen Lba Cas12a (Cpf1) - 70 pmol $99.00
$89.10
ADD TO CART
M0653T EnGen Lba Cas12a (Cpf1) - 2,000 pmol $355.00
$319.50
ADD TO CART
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

    Product Introduction

    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' TTTN protospacer adjacent motif (PAM) on the DNA strand opposite the target sequence. Cleavage by EnGen Lba Cas12a (Cpf1) occurs ~18 bases 3' of the PAM and leaves 5' overhanging ends.

    • T-rich TTTN 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

    Product Information

    Description

    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’ TTTN 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:

    Store at (°C) Concentration
    NEBuffer™ 2.1 -20 10X
    Product Categories:
    Genome Editing Products
    Applications:
    Genome Editing Applications

    Properties & Usage

      Reaction Conditions

      1X NEBuffer™ 2.1
      Incubate at 37°C

      1X NEBuffer™ 2.1
      50 mM NaCl
      10 mM Tris-HCl
      10 mM MgCl2
      100 µg/ml BSA
      (pH 7.9 @ 25°C)

      Storage Temperature

      -20°C

      Storage Conditions

      500 mM NaCl
      20 mM sodium acetate
      0.1 mM EDTA
      0.1 mM TCEP
      50% Glycerol
      pH 6.0 @ 25°C

      Heat Inactivation

      65°C for 10 min

      Product Notes

      1. 100 µM is equal to 15.1 mg/ml EnGen Lba Cas12a.

      References

      1. Zetsche, B., et. al. (2015). Cell. 163, 759-771.
      2. Moreno-Mateos, M.A., et al. (2017). Nat. Commun.. 8, 2024. PubMedID: 29222508,

      Protocols, Manuals & Usage

      Protocols

      1. Protocol for Electroporation of Cas12a Ribonucleoprotein (RNP) into adherent cells using the Neon® Electroporation
      2. Protocol for Electroporation of Cas12a Ribonucleoprotein (RNP) into adherent cells using the Lonza 4D-Nucleofector®
      3. In vitro digestion of DNA with EnGen® Lba Cas12a (Cpf1) (M0653)

      Faqs & Troubleshooting

      FAQs

      1. What are the differences between EnGen® Lba Cas12a (Cpf1) (NEB #M0653) and EnGen Spy Cas9 NLS (NEB #M0646)?
      2. Which nuclear localization signal is fused to EnGen® Lba Cas12a (Cpf1)?
      3. Why am I observing incomplete digestion with EnGen® Lba Cas12a (Cpf1)?
      4. Why does digestion efficiency differ between two different gRNAs?
      5. Can mutations generated with EnGen Lba Cas12a (Cpf1) be detected using T7 Endonuclease I (NEB #M0302) or the EnGen Mutation Detection Kit (NEB #E3321S)?
      6. Can gRNA for use with EnGen Lba Cas12a (Cpf1) be generated using the EnGen sgRNA Synthesis Kit, S. pyogenes (NEB #E3322)?
      7. How do I design a guide RNA for use with EnGen Lba Cas12a?

      Troubleshooting

      Quality, Safety & Legal

      Quality Assurance Statement

      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 DataSheets

      The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

      Legal and Disclaimers

      This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

      While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

      For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

      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.