LbCas12a is an RNA-guided endonuclease that is naturally expressed by Lachnospiraceae bacterium. The recombinant enzyme is engineered as a fusion protein with a eukaryotic nuclear localization signal (NLS) at the C-terminus to promote import of LbCas12a/gRNA ribonucleoprotein (RNP) complexes into the mammalian cell nucleus for genome engineering purposes.

Presence of the NLS does not affect the biochemical activity of LbCas12a when used for in vitro applications. Targeting of LbCas12a requires complexing of the enzyme to a guide RNA (gRNA) in a 1:1 ratio to form an active RNP complex. The gRNA is composed of a common 20-nucleotide direct repeat (DR) sequence followed by a variable 21-nucleotide guide sequence that is complementary to the DNA target strand.

The guide sequence must be complementary to target site with a 5' TTTV protospacer adjacent motif (PAM) on the DNA strand opposite the target sequence. Upon recognition of its DNA target sequence, LbCas12a will cleave both strands about 17 bases 3' to the PAM site and leave 5' overhangs on both strands.

Store at −20 °C upon arrival until provided expiration date. See individual component labels for additional storage recommendations. Minimize freeze/thaw cycles to avoid loss of performance. Maintain product on ice during use.

Storage buffer: LbCas12a cleavage buffer: 10X