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現(xiàn)貨促銷(xiāo)CRISPR /Cas9專(zhuān)家系統(tǒng)服務(wù)-ARCA Cas9 mRNA/sgRNA

2023-04-05

品牌: biomics 公司: 百奧邁科生物技術(shù)有限公司

提供商: Biomics Biotech 服務(wù)名稱(chēng): ARCA Cas9 mRNA/sgRNA 規(guī)格: 0.5-1μg/μL

產(chǎn)品詳情請(qǐng)聯(lián)系金山科研平臺(tái)微信:jinshanbio
近10年來(lái),mRNA在醫(yī)學(xué)應(yīng)用領(lǐng)域取得顯著進(jìn)展。研究人員可快速合成任意序列的mRNA片段,并能保存在室溫條件下。在無(wú)細(xì)胞的體系中利用DNA轉(zhuǎn)錄生成cap和poly A修飾的mRNA分子穩(wěn)定性良好,而修飾堿基的mRNA在穩(wěn)定性上更獲得大幅提升。在研究基因敲除動(dòng)物及其胚胎發(fā)育中的基因表達(dá)時(shí),科研人員往往都采用卵母細(xì)胞的mRNA顯微注射的方法,該方法在ZFN、TALEN、CRISPR、重編程細(xì)胞多能性等技術(shù)中得到廣泛應(yīng)用。在這些基因編輯過(guò)程中使用mRNA較之DNA有諸多優(yōu)點(diǎn),例如無(wú)需進(jìn)入核內(nèi)并轉(zhuǎn)錄效率更快,RNA易降解也避免基因組遺傳重組現(xiàn)象。鑒于此,Biomics Biotech推出mRNA系列產(chǎn)品,提供不同類(lèi)型的Cas9 mRNA, 例如野生型spCas9 mRNA、單突變spCas9 Nickase mRNA、雙突變spCas9 mutant mRNA。此外我們還提供各種報(bào)告基因的mRNA給客戶作為陽(yáng)性對(duì)照,例如Luciferase mRNA,mCHERRY mRNA, GFP mRNA, β-gal mRNA。Biomics Cas9 mRNA 使用本公司自主開(kāi)發(fā)的T7 cap mRNA MICscript? KIT轉(zhuǎn)錄合成和EzOmicsTM RNA Quick Clear Kit純化,從而使我們的產(chǎn)品成本更低,較客戶購(gòu)買(mǎi)國(guó)外同類(lèi)產(chǎn)品更具性?xún)r(jià)比。Cas9 mRNA轉(zhuǎn)錄DNA模板為線性化質(zhì)粒,確保無(wú)冗余轉(zhuǎn)錄序列。該質(zhì)粒中Cas9基因?yàn)槿嗽疵艽a子優(yōu)化spCas9基因,帶有核轉(zhuǎn)導(dǎo)域(NLS)及Poly A信號(hào)(globin3’ UTR),Cas9 mRNA在細(xì)胞內(nèi)更具穩(wěn)定性。細(xì)胞注射或轉(zhuǎn)染用mRNA合成還有一個(gè)關(guān)鍵步驟是5’端的加帽,帽子結(jié)構(gòu)是指在真核生物中轉(zhuǎn)錄后修飾形成的成熟mRNA在5'端的一個(gè)特殊結(jié)構(gòu),即m7GPPPN結(jié)構(gòu),又稱(chēng)為甲基鳥(niǎo)苷帽子。它是在RNA三磷酸酶,mRNA鳥(niǎo)苷酰轉(zhuǎn)移酶,mRNA(鳥(niǎo)嘌呤-7)甲基轉(zhuǎn)移酶和mRNA(核苷-2’)甲基轉(zhuǎn)移酶催化形成的。mRNA 5’-端帽子結(jié)構(gòu)是mRNA翻譯起始的必要結(jié)構(gòu),對(duì)核糖體對(duì)mRNA的識(shí)別提供了信號(hào),協(xié)助核糖體與mRNA結(jié)合,使翻譯從AUG開(kāi)始。帽子結(jié)構(gòu)可增加mRNA的穩(wěn)定性,保護(hù)mRNA免遭5’ →3‘核酸外切酶的攻擊。Biomics使用抗反向帽子類(lèi)似物(ARCA) m27,3'-OG[5']ppp[5']在體外產(chǎn)生加帽RNA。在體外的轉(zhuǎn)錄反應(yīng)中由于ACRA的m7G核苷上含有一個(gè)3’-O甲基,ACRA可僅被整合到RNA5‘端’正確的方向。因此,ARCA的整合導(dǎo)致加帽RNA的合成比標(biāo)準(zhǔn)帽子類(lèi)似物(mCAP、CAP)翻譯更加有效。 Biomics mRNA產(chǎn)品濃度為0.5-1μg/μL ,文獻(xiàn)報(bào)道中顯微注射的mRNA使用濃度一般為20-200ng/μL 關(guān)聯(lián)產(chǎn)品:
    T7 cap mRNA MICscript? KITEzOmicsTM RNA Quick Clear KitReporter mRNACustom Long RNA Synthesis ServicesReprogramming mRNA
FAQ:
    In eukaryotes: the 3’ poly(A) tail confers stability.
A: Most mammalian mRNAs are Polyadenylated
    mRNA Decay and Translation
A: The intimate relationship between mRNA decay and translation is further indicated by the ability of translation-initiation factors (eIF) and proteins (PAB) that bind the poly (A) tail to protect the mRNA from degradation. Moreover, evidence shows that inhibiting translation elongation promotes mRNA stabilization.Translation initiation complex
    What is the rate-limiting step in mRNA degradation?
A: An evolutionarily conserved mRNA-degradation pathway is initiated by the removal of the 3’- poly(A) tail. This disrupts the translation initiation complex and provides degradative enzymes with access to the 5’ cap and remaining RNA body.Reference:
    McIvor RS. Therapeutic Delivery of mRNA: The Medium Is the Message,Mol Ther. 2011 May; 19(5):822-3.Warren et al., Highly Efficient Reprogramming to Pluripotency and Directed Differentiation of Human Cells with Synthetic Modified mRNA, Cell Stem Cell (2010), Yang H, Wang H, Shivalila CS, Cheng AW, Shi L, Jaenisch R. One-step generation of mice carrying reporter and conditional alleles by CRISPR/Cas-mediated genome engineering. Cell. 2013 Sep 12; 154(6):1370-9.Chang N1, Sun C, Gao L, Zhu D, Xu X, Zhu X, Xiong JW, Xi JJ. Genome editing with RNA-guided Cas9 nuclease in Zabrafish embryos. Cell Res. 2013 Apr; 23(4):465-72.Ma Y, Shen B, Zhang X, Lu Y, Chen W, Ma J, Huang X, Zhang L. Heritable multiplex genetic engineering in rats using CRISPR/Cas9. PLoS One. 2014 Mar 5; 9(3):e89413.Sung YH, Kim JM,Kim HT,Lee J, Highly efficient gene knockout in mice and Zabrafish with RNA-guided endonuclease. Genome Res. 2014 Jan; 24(1):125-31.
CRISPR/Cas9技術(shù)討論群:342716231微信號(hào):Biomics-RNAi

資料下載:

中文版 Biomics ARCA Cas9 mRNA manual.pdf 附件下載 (下載 6 次)

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