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.
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.
N-type Ca CP α1B双切酶质粒(h)和N-type Ca CP α1B双切酶质粒(h2)编码针对CACNA1B的不同配对gRNA设计。其中一种或两种设计可能均有提供
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N-type Ca CP α1B双切口酶质粒(h)
sc-402397-NIC
20 µg
CNY3084.00
N-type Ca CP α1B双切口酶质粒(h2)
sc-402397-NIC-2
20 µg
CNY3084.00
CACNA1B 编码 N 型电压门控钙通道(CaV2.2)的成孔 α1B 亚基,介导兴奋性细胞在去极化刺激下的 Ca2+ 内流。该通道将膜电活动与突触前囊泡融合及神经递质释放相耦联,并整合进 Ca2+ 依赖的信号网络,从而调控突触可塑性、神经元兴奋性以及活动依赖性的基因表达。CACNA1B 的功能与 G 蛋白偶联受体及其下游通路对突触传递动力学的调制密切相关。遗传变异或 CaV2.2 活性失调与伤害感受处理的改变及神经精神表型有关,提示其在解析神经回路功能机制的研究中具有重要意义。
N-type Ca CP α1B 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 CACNA1B 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CACNA1B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CACNA1B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。