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.

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Schematic representation of Lba Cas12a nuclease sequence recognition and DNA cleavage

Expanded temperature range of Lba Cas12a

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:

Show all

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

M0653S

?

?

-20

?

?

?

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

M0653T

?

?

-20

?

?

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