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 cleavageExpanded temperature range of Lba Cas12aPurified 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.
This product is related to the following categories:
Genome Editing Products,
Programmable Nucleases
This product can be used in the following applications: