Date published: 2026-7-17
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The Double Nickase Plasmid features a U6 promoter for sgRNA expression, a 20 nt targeting sequence, and a gRNA scaffold to guide Cas9n. It includes a CBh promoter for Cas9n (D10A) and puromycin resistance, GFP for transfection verification, and nuclear localization signals (NLS). The 2A peptide allows co-expression of Cas9n and Puro from a single promoter, enabling precise genome editing with reduced off-target effects.
The Double Nickase Plasmid features a U6 promoter for sgRNA expression, a 20 nt targeting sequence, and a gRNA scaffold to guide Cas9n. It includes a CBh promoter for Cas9n (D10A) and puromycin resistance, GFP for transfection verification, and nuclear localization signals (NLS). The 2A peptide allows co-expression of Cas9n and Puro from a single promoter, enabling precise genome editing with reduced off-target effects.
The Double Nickase Plasmid features a U6 promoter for sgRNA expression, a 20 nt targeting sequence, and a gRNA scaffold to guide Cas9n. It includes a CBh promoter for Cas9n (D10A) and puromycin resistance, GFP for transfection verification, and nuclear localization signals (NLS). The 2A peptide allows co-expression of Cas9n and Puro from a single promoter, enabling precise genome editing with reduced off-target effects.
The Double Nickase Plasmid features a U6 promoter for sgRNA expression, a 20 nt targeting sequence, and a gRNA scaffold to guide Cas9n. It includes a CBh promoter for Cas9n (D10A) and puromycin resistance, GFP for transfection verification, and nuclear localization signals (NLS). The 2A peptide allows co-expression of Cas9n and Puro from a single promoter, enabling precise genome editing with reduced off-target effects.
Cas9n Nickase gRNA Plasmid Targeting: Dual gRNA plasmids create single-strand nicks at precise DNA sequences for efficient genome editing using Cas9n Nickase.
Cas9n Nickase gRNA Plasmid Targeting: Dual gRNA plasmids create single-strand nicks at precise DNA sequences for efficient genome editing using Cas9n Nickase.
This image illustrates the Cas9n Nickase mechanism used for precise genome editing. Two plasmids (Plasmid 1 and Plasmid 2) are shown, each containing a targeted DNA sequence. The system utilizes single-guide RNAs (sgRNA) to direct Cas9n Nickase to specific genomic locations, represented by the blue and pink DNA strands. The sgRNA scaffold aids in guiding Cas9n to the 20 nucleotide (nt) target sequence on the DNA. Cas9n makes single-strand cuts at NCC and NGG sites, enabling precise gene modifications without creating double-strand breaks.
This image illustrates the Cas9n Nickase mechanism used for precise genome editing. Two plasmids (Plasmid 1 and Plasmid 2) are shown, each containing a targeted DNA sequence. The system utilizes single-guide RNAs (sgRNA) to direct Cas9n Nickase to specific genomic locations, represented by the blue and pink DNA strands. The sgRNA scaffold aids in guiding Cas9n to the 20 nucleotide (nt) target sequence on the DNA. Cas9n makes single-strand cuts at NCC and NGG sites, enabling precise gene modifications without creating double-strand breaks.
The Double Nickase Plasmid features a U6 promoter for sgRNA expression, a 20 nt targeting sequence, and a gRNA scaffold to guide Cas9n. It includes a CBh promoter for Cas9n (D10A) and puromycin resistance, GFP for transfection verification, and nuclear localization signals (NLS). The 2A peptide allows co-expression of Cas9n and Puro from a single promoter, enabling precise genome editing with reduced off-target effects.

NFκB p50双切口酶质粒(h): sc-400087-NIC

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说明书
  • 针对种属：human
  • 20 µg 纯化的即用型的质粒DNA，最多可供20次转染
  • NFκB p50 双切口酶质粒（h）含有一对质粒，分别编码D10A突变的Cas9核酸酶，和目标特异的 20 nt 向导RNA （gRNA），与其相应的/%base_sku_name%/ CRISPR/Cas9敲除质粒相比，它在基因敲除方面具有更好的特异性
  • 成对的向导RNA序列与目标基因中约20个碱基对互补，从而使Cas9可以模仿DNA双链断裂（DSB），介导特异的基因组DNA双切割
  • 质粒对中的一个包含供筛选使用的嘌呤霉素抗性基因；另一个包含供观察转染使用的绿色荧光蛋白标记
  • NFκB p50双切酶质粒（h）和NFκB p50双切酶质粒（h2）编码针对NFKB1的不同配对gRNA设计。其中一种或两种设计可能均有提供
  • 转染后，基因敲除效率可以用抗体：NFκB p50: sc-8414，通过WB, IF或者IHC分析
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    订购信息

    产品名称产品编号规格价格数量收藏夹

    NFκB p50双切口酶质粒(h)

    sc-400087-NIC
    20 µg
    		$410.00

    NFκB p50双切口酶质粒(h2)

    sc-400087-NIC-2
    20 µg
    		$410.00

    NFKB1 编码 NFκB 的 p50 亚基，它是 NF-κB 转录因子复合物的核心组成部分。这些复合物在炎性细胞因子刺激、Toll 样受体信号传导、抗原受体激活以及细胞应激等条件下，调控可诱导基因的表达。NFκB p50 由 p105 前体经蛋白水解加工产生，可形成同源或异源二聚体（常与 RelA/p65 配对），从而控制与先天与适应性免疫、细胞存活及分化相关的转录程序。通过经典与非经典 NF-κB 通路，p50 还会影响染色质可及性以及细胞因子/趋化因子网络，从而塑造机体对感染和组织损伤的应答。NFKB1 活性失调，以及其与 MAPK、JAK/STAT 和干扰素信号之间的通路串扰，常被认为与慢性炎症、免疫功能紊乱以及促肿瘤的微环境密切相关。

    NFκB p50 双切酶质粒（h）由一对匹配的质粒组成，专为在 human 细胞系中对 NFKB1 位点进行高特异性编辑而设计。每个质粒分别表达Cas9 D10A切口酶和针对NFKB1内不同DNA链的独特sgRNA。当这两种切口酶被引导至相邻但位于DNA链相反侧的位点时，会产生错位的单链切口，从而共同形成错位双链断裂，这需要两个引导RNA在靶位点上协同发挥作用。由此产生的DNA断裂通过内源性细胞修复途径（最常见的是非同源末端连接（NHEJ））得到修复，从而导致插入或缺失，进而破坏NFKB1的功能。通过要求双sgRNA在靶位点结合，双切口方法提高了编辑特异性，并为需要对靶向精度进行额外控制的应用提供了互补的CRISPR策略。

    为高效识别编辑后的细胞，其中一个质粒编码GFP以实现转染细胞群的荧光可视化，而配套质粒则携带嘌呤霉素抗性基因用于抗生素筛选。这些特性共同支持共转染细胞群的高效富集，并简化了NFKB1基因失活克隆的验证流程。

    仅供研究使用。不用于诊断或治疗。

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