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產品名稱:pCas9(ZK1672)

貨號 規格 價格 訂購數量 是否現貨
ZK1672 1ug×1支 580 - + 有貨
質粒分類
主要功能次級功能用途抗性
大腸桿菌質粒
大腸編輯質粒
大腸Cas9質粒
基因敲除
Chl


■ 基本信息

復制子: p15A ori

質粒分類: 大腸系列質粒;大腸編輯質粒;大腸Cas9質粒

質粒大小: 9326bp

原核抗性: 氯霉素Chl(50μg/ml)

克隆菌株: Stbl3等大腸桿菌

培養條件: 37℃,LB,有氧

表達宿主: 大腸桿菌

誘導方式: 無需誘導

5'測序引物: StrepPyoCas9-5UTR-F(CGGTGCCACTTTTTCAAGTTG)

3'測序引物: pBRrevBam(GGTGATGTCGGCGATATAGG)


■ 質粒簡介

pCas9質粒是CRISPR/Cas9系統類載體,復制子是p15A ori和ori,質粒大小是9326bp,可轉化進相應的宿主細胞中,通大量養條件是LB,37℃。

pCas9可在大腸桿菌中表達Cas9蛋白,和pCRISPR搭配可用于基因敲除,低拷貝質粒,Bacterial expression of Cas9 nuclease, tracrRNA and crRNA guide.

pCas9 can often be difficult to target double-stranded breaks (DSBs) with the precision that is necessary for various genome editing applications. The ability to engineer Cas9 derivatives with purposefully altered PAM specificities would address this limitation. Here we show that the commonly used Streptococcus pyogenes Cas9 (SpCas9) can be modified to recognize alternative PAM sequences using structural information, bacterial selection-baseddirected evolution, and combinatorial design. CRISPR-Cas9 nucleases enable efficient genome editing in a wide variety of organisms and cell types . Target site recognition by Cas9 is programmed by a chimeric single guide RNA (sgRNA) that encodes a sequence complementary to a target protospacer5 , but also requires recognition of a short neighboring PAM . SpCas9, the most robust and widely used Cas9 to date, primarily recognizes NGG PAMs and is consequently restricted to sites that contain this motif . It can therefore be challenging to implement genome editing applications that require precision, such as: homology-directed repair (HDR), which is most efficient when DSBs are placed within 10-20 bps of a desired alteration; the introduction of variable-length insertion or deletion (indel) mutations into small size genetic elements such as microRNAs, splice sites, short open reading frames, or transcription factor binding sites by non-homologous end-joining (NHEJ); and allele-specific editing, where PAM recognition might be exploited to differentiate alleles.


載體質粒圖譜和多克隆位點信息


ZK1672 pCas9大腸質粒說明書.doc


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