AAVS1 sgRNA Cas9靶向腺相关病毒整合位点1的CRISPR-Cas9 基因编辑质粒载体-BIoVector保藏中心

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货　　号：BioVector-AAVS1 sgRNA Cas9
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BIoVector™ AAVS1 sgRNA Cas9 plasmid is a specialized tool used for CRISPR-Cas9 genome editing that simplifies the process of integrating a gene into a specific, safe location in the human genome. It combines the three core components of the CRISPR system—the Cas9 nuclease, the single guide RNA (sgRNA), and the plasmid backbone—into a single vector.

What makes this plasmid unique is that its sgRNA is specifically designed to target the AAVS1 locus. The AAVS1 (Adeno-Associated Virus Integration Site 1) locus, located on human chromosome 19, is known as a "genomic safe harbor". This means that a gene inserted here is highly likely to be expressed stably and at high levels without causing harm to the cell or disrupting the function of other genes.

All-in-one Cas9 SmartNuclease™ & AAVS1 gRNA Plasmid | System Biosciences

Components and Function

The plasmid contains several key elements:

Cas9 gene: This codes for the Cas9 endonuclease, the "molecular scissors" that cuts DNA.
AAVS1 sgRNA: This is a short RNA molecule that acts as a guide. It directs the Cas9 protein to the specific AAVS1 locus on chromosome 19.
Plasmid backbone: This is the circular DNA molecule that carries all the components and allows for replication within bacteria.

All-in-one Cas9 SmartNuclease™ & AAVS1 gRNA Plasmid

When this "all-in-one" plasmid is introduced into human cells, the sgRNA guides the Cas9 protein to the AAVS1 site, where it creates a double-strand break. Researchers can then introduce a separate donor plasmid containing their gene of interest. The cell's natural DNA repair mechanism, called homology-directed repair (HDR), will use the donor plasmid as a template to insert the new gene precisely into the AAVS1 locus.

Benefits and Applications

Using an AAVS1 sgRNA Cas9 plasmid offers several major advantages over traditional methods of gene integration:

Stable and Predictable Expression: Integration into a safe harbor site ensures that the inserted gene remains active and is not silenced over time, a common issue with random integration.
Reduced Off-Target Effects: By targeting a known, well-characterized site, the risk of disrupting a critical gene or causing an adverse effect is minimized.
Simplified Cell Line Generation: It streamlines the process of creating stable, isogenic (genetically uniform) cell lines for research, such as creating disease models or reporter cell lines.
Precise Knock-in: This system is ideal for precise gene knock-in, a process where a new gene is inserted into a specific location in the genome.

This type of plasmid is widely used in applications like creating cell-based disease models, developing transgenic cell lines, and performing functional genomics screens.

AAVS1 sgRNA Cas9 质粒 是一种专门用于靶向腺相关病毒整合位点1的CRISPR-Cas9 基因编辑的工具，它简化了将特定基因整合到人类基因组中一个安全且稳定的位置的过程。它将 CRISPR 系统的三个核心组件——Cas9 核酸酶、单向导 RNA（sgRNA）和质粒骨架——组合在一个载体中。

它的独特之处

这种质粒的独特之处在于，它的 sgRNA 专门设计用于靶向 AAVS1 基因座。AAVS1（腺相关病毒整合位点 1）基因座位于人类 19 号染色体上，它被称为一个 “基因组安全港”。这意味着，如果一个基因被插入到这个位置，它极有可能稳定且高水平地表达，同时不会对细胞造成伤害或扰乱其他基因的功能。

质粒的组成与功能

该质粒包含几个关键元件：

Cas9 基因： 编码 Cas9 核酸内切酶，即 DNA 的“分子剪刀”，负责切割 DNA。
AAVS1 sgRNA： 这是一种短的 RNA 分子，充当向导。它将 Cas9 蛋白引导至 19 号染色体上特定的 AAVS1 基因座。
质粒骨架： 这是一个环状 DNA 分子，携带所有元件，并允许在细菌中复制。

当这种“多合一”质粒被导入人类细胞后，sgRNA 会引导 Cas9 蛋白到达 AAVS1 位点，并在那里创建一个双链断裂。随后，研究人员可以导入一个单独的供体质粒，其中包含他们感兴趣的基因。细胞自身的 DNA 修复机制，称为同源定向修复（HDR），会利用供体质粒作为模板，将新基因精确地插入到 AAVS1 基因座。