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:
核糖核酸酶A是内切核糖核酸酶,可特异地攻击RNA上嘧啶残基的3'端,切割与相邻核苷酸形成的磷酸二酯键。反应终产物是嘧啶3'磷酸及末端带嘧啶3'磷酸的寡核苷酸。无辅因子及二价阳离子存在时,核糖核酸酶A的作用可被胎盘RNA酶抑制剂(B1ackburn et al.1977)或氧钒—核糖核苷复合物(Puskas et al.1982)所抑制。核糖核酸酶A用途生化研究,测定核酸的结构RNase 保护检测去除非... 查看更多>
DNase I 是一种核酸内切酶,降解双链或单链DNA,产生5-磷酸末端的单核苷酸及寡核苷酸,在Mg2+存在时,DNase I独立地作用每条DNA链,切割位点是随机分布,在Mn2+存在下,DNase I 作用于DNA双链的大致同一位置,产生钝末端或具1-2个核苷酸突起的DNA片段。来源:牛胰贮存条件:4℃应用:1、用切口平移法进行放射性标记时,可用DNase I在双链DNA上产生随机切口;2、在进行亚硫酸氢盐介导的诱变前,可用DNase... 查看更多>