一般描述
CRISPR植物Cas9产品可用于农杆菌属介导的植物转化或基因枪微粒轰击或原生质体转化。该产品是基于酿脓链球菌的IIA型CRISPR-Cas9。天然的Cas9编码序列经过密码子优化，可分别用于在单子叶植物和双子叶植物中进行表达。单子叶植物Cas9构建体含有可驱动sgRNA表达的单子叶植物U6启动子，双子叶植物Cas9构建体含有双子叶植物U6启动子。
植物筛选标记物包括：
- 潮霉素B抗性基因
- 新霉素磷酸转移酶基因
- 抗除草剂基因（草丁膦乙酰转移酶）
应用
- 基因失活
- 靶标验证
- 目的基因的位点特异性整合
- 通过HR替换基因
特点和优势
- CRISPR/Cas9的主要优势在于其具有简单性、易用性、低成本和通用性。
- CRISPR/Cas9系统不需要任何蛋白质工程步骤，更易于检测每个目标基因的多个gRNA
- 只需改变gRNA序列中的20个核苷酸，即可获得不同的靶点特异性
- 与ZFN和TALEN相比，CRISPR/Cas9的另一个优势在于它可同时在多个位点导入DSB，从而实现对多个基因同时进行编辑。这对于敲除多余基因或平行通路特别有帮助。使用CRISPR/Cas9系统进行同时编辑，只需单体Cas9蛋白以及任何数量的、不同的序列特异性gRNA。
- CRISPR/Cas9系统可切割人类细胞中的甲基化DNA，其达到的基因修饰超越了其他核酸酶。这一点尚未在植物中进行探索，可以认为切割甲基化DNA是CRISPR/Cas9系统特有的功能，并且与目标基因组无关。
组分
在不使用时，将试管盖扣紧。
利用无菌实验技术，避免DNAase污染。
原理
其他说明
法律信息
储存分类代码
12 - Non Combustible Liquids
WGK
WGK 1
闪点(°F)
Not applicable
闪点(°C)
Not applicable
法规信息
分析证书(COA)
输入产品批号来搜索 分析证书(COA) 。批号可以在产品标签上"批“ （Lot或Batch）字后找到。
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示例
其它示例：
705578-5MG-PW
PL860-CGA/SHF-1EA
MMYOMAG-74K-13
1000309185
输入内容 1.000309185)
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Aldrich 产品
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If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
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1. What type of Cas9 is used in the CRISPR Plant vectors?
Type IIA Cas9 derived from Streptococcus pyogenes.
How is the Cas9 codon sequence optimized?
In the monocot CRISPR Plant vectors, the Cas9 codon optimization is based on Zea mays. In the dicot CRISPR Plant vectors, the Cas9 codon optimization is based on Arabidopsis thaliana.
What is the promoter used for the expression of Cas9?
The promoter used is 2X35S, derived from cauliflower mosaic virus (CaMV).
What is the promoter used for the expression of the selection markers?
The 2X35S promoter is used in this case.
What is the promoter used for sgRNA (single guide RNA) expression?
In the monocot CRISPR vectors, it is a wheat U6 promoter. In the dicot CRISPR vectors, it is an Arabidopsis thaliana U6 promoter.
Can one use an Agrobacterium-mediated transformation CRISPR vector for biolistics or protoplast plant transformation?
Yes. However, the biolistics/protoplast CRISPR vectors cannot be used for Agrobacterium-mediated plant transformation.
What antibiotics should I use for selection of the CRISPR Plant vectors in E. coli or Agrobacteria tumefaciens?
Kanamycin.
Can I use the monocot CRISPR plant vectors on dicots plants and the dicot CRISPR plant vectors on monocot plants?
Yes. But it is not recommended. The monocot CRISPR plant vectors are more suited for monocot plants, and the dicots CRISPR plant vectors are more suited for dicot plants.
Why did my CRISPR plant construct did not produce the expected target cleavage?
Not every sgRNA will produce the target cleavage. It is recommended to use two to three sgRNAs per gene to increase the successful rate.
How can I use the CRISPR plant vectors for homologous recombination (HR)?
First, you need to design an HR DNA donor with the target of interest and the homologous arms. For Agrobacterium-mediated plant transformation, the HR DNA donor can be inserted into an appropriate restriction site, such as the EcoRI site or the HindIII site, within the T-DNA borders. For biolistics or protoplast plant transformation, the HR DNA donor can be inserted into XbaI, EcoRI, HindIII, or XhoI site. Alternatively the HR DNA donor can be co-delivered in a separate donor vector.
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