Date published: 2026-7-12
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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.

GCK双切口酶质粒(h): sc-400852-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
  • GCK 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
  • GCK双切酶质粒（h）和GCK双切酶质粒（h2）编码针对GCK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：GCK: sc-17819，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

    GCK双切口酶质粒(h)

    sc-400852-NIC
    20 µg
    		$410.00

    GCK双切口酶质粒(h2)

    sc-400852-NIC-2
    20 µg
    		$410.00

    葡萄糖激酶（GCK）编码己糖激酶IV型酶，可将葡萄糖磷酸化生成葡萄糖-6-磷酸，作为关键的葡萄糖传感器，将细胞外葡萄糖的可获得性与细胞代谢相耦联。在胰岛β细胞中，GCK活性有助于设定葡萄糖刺激胰岛素分泌的阈值；而在肝细胞中，GCK通过与糖酵解、糖异生调控以及碳水化合物响应性代谢程序的整合，支持糖酵解通量和糖原合成。通过控制葡萄糖进入中心碳代谢，GCK影响ATP/ADP平衡、代谢物信号传导以及与胰岛素分泌和肝脏葡萄糖利用相关的下游通路。GCK的遗传或功能性扰动与葡萄糖稳态异常及单基因型血糖异常密切相关，因此它也是研究代谢调控机制时广泛使用的靶点。

    GCK 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 GCK 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GCK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GCK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GCK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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