pYLCRISPR/Cas9P35S-N植物多基因座基因组编辑二元载体质粒BioVector® NTCC保藏中心

BioVector® pYLCRISPR/Cas9P35S-N 是一种用于植物多基因座基因组编辑的二元载体（Binary Vector），特别适用于双子叶和单子叶植物。

• 载体类型： 植物 CRISPR/Cas9 表达二元载体

• 细菌抗性： 卡那霉素 (Kanamycin, 50 μg/mL)

• 植物筛选标记：NPTII (赋予植物对 G418 或卡那霉素的抗性)

• Cas9 启动子： 使用 Cauliflower Mosaic Virus 35S (P35S) 启动子，在大多数植物组织中呈强组成型表达。

• Cas9 基因： 采用经过植物密码子优化（Plant-codon optimized）的 S. pyogenes Cas9，提高了在植物细胞中的翻译效率。

• 多靶点构建： 该载体属于 pYLCRISPR 系列，支持通过 Golden Gate 组装（BsaI 位点）同时插入多个 sgRNA 表达盒。

• 适用植物： 广泛应用于拟南芥、烟草以及多种园艺作物。

• 转化方式： 通常通过农杆菌介导法（Agrobacterium-mediated transformation）转化植物。

General Specifications

• Vector Type: Plant binary vector for CRISPR/Cas9 expression.

• Bacterial Resistance: Kanamycin (50 μg/mL).

• Plant Selection Marker: Neomycin (NPTII), conferring resistance to G418/Kanamycin in plants.

• Cas9 Expression: Driven by the Cauliflower Mosaic Virus 35S promoter (P35S), which provides strong constitutive expression in most plant tissues.

• Cas9 Gene/Insert:S. pyogenes Cas9, plant-codon optimized with higher GC content (62.5% in the 5' region; 54.2% overall) to enhance translation efficiency in plants.

• Multiplexing Capability: Compatible with the pYLCRISPR system for assembling multiple sgRNA expression cassettes (driven by various promoters like AtU3b, AtU3d, AtU6-1, or AtU6-29) using Golden Gate ligation.

• Primary Reference:Ma et al. (2015), "A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants".

• Common Hosts:Arabidopsis, tobacco (N. benthamiana), and other dicots; also used in various horticultural crops and perennial plants.

• Transformation: Typically mediated by Agrobacterium tumefaciens (e.g., strains EHA105 or GV3101).