Each CRISPR/Cas9 Knockout (KO) Plasmid product consists of a pool of 3 plasmids designed to ensure identification and cleavage of a specific gene for maximum knockout efficiency. Each plasmid encodes a unique 20 nt sequence derived from the GeCKO (v2) library.
Each CRISPR/Cas9 Knockout (KO) Plasmid product consists of a pool of 3 plasmids designed to ensure identification and cleavage of a specific gene for maximum knockout efficiency. Each plasmid encodes a unique 20 nt sequence derived from the GeCKO (v2) library.
All products are provided as transfection-ready, purified plasmid DNA, except the Lentiviral Activation Particles which have been prepackaged as Lentiviral Particles for use with hard-to-transfect cells
CRISPR/Cas9 KO plasmids, Double Nickase Plasmids and CRISPR/dCas9 Activation plasmids are considered a "Licensed Product" and are to be used in accordance with the Limited/Label License. Click here for details
Description
20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
LAT Double Nickase Plasmid (m) 는 한쌍의D10A mutated Cas9 nuclease 와 타깃특정 20 nt guide RNA (gRNA)를 인코딩하는 plasmid를 포함하고있으며 gRNA는 대응 CRISPR/Cas9 KO보다 높은 특이성을 띤 knockout gene을 발현하도록 디자인되였습니다.
한쌍의 gRNA는 약 20bp가량 떨어져 있으며 이는 게놈DNA의 Cas9-mediated double nicking을 발생하여 DSB를 모방합니다.
한쌍의 plasmid 중 하나는 selection에 필요한 puromycin-resistance gene을, 다른하나는 형질주입효율을 확인하는 GFP marker를 보유하고있습니다.
LAT Double Nickase Plasmid (m2)가 인코딩한 한쌍의 gRNAs는 LAT Double Nickase Plasmid (m)가 인코딩한것과 다릅니다.
Transfection, gene knockout효율은 WB, IF나 IHC등 방법으로 측정할수있습니다, 권장하는 항체는 LAT Antibody (11B.12): sc-53550