siRNA pools（siPOOLs）

一款高效、特異性的RNAi產品

對于靶向基因沉默，RNA干擾（RNAi)具有易于操作、快速結果、高效率、廣泛適用于各種細胞類型的多重優(yōu) 勢。其具有瞬轉效應和劑量依賴效應，這點與小分子非常相似。然而，目前的siRNA試劑其特異性和基因沉默 效率可變，阻礙了它們作為藥物發(fā)現(xiàn)和基因研究工具的應用。

siTOOLs Biotech由Michael Hannus博士和Gunter Meister教授于2013年9月成立，由德國雷根斯堡大學 和Intana Bioscience制藥服務公司聯(lián)合運營。目前推出有3個產品系列：（1）siPOOLs：一種siRNA混合物，有 效增加基因沉默特異性，“稀釋”傳統(tǒng)RNA干擾試劑的脫靶效應。（2）raPOOLs：一種穩(wěn)健的RNA親和純化方 案，適用于基因功能和相互作用的生物化學研究。（3）riboPOOLs：適合任意物種的核糖體RNA去除試劑，高效且經濟。

可靠表型的特異性基因沉默工具

siPOOLs是一款經過優(yōu)化設計的高復雜性的包含30條siRNA的混合物，經證明可有效消除脫靶效應，并提高結果的可靠性（Hannus et al., 2014）。 Pack Hunter (pooling) 方法通過將當個siRNA的 濃度稀釋到刺激表型的閾值以下來對抗單個siRNA的脫靶。 借助專有的設計算法，siPOOLs中的siRNA序列經 過優(yōu)化，以實現(xiàn)最大的轉錄本覆蓋率，高效雜交并 對旁系同源基因進行過濾，從而實現(xiàn)高效和特異性 的基因沉默。

siPOOLs產品優(yōu)勢

使用簡單快捷：siPOOLs與多種轉染試劑兼容，幾天內就能看 到結果。 

高度特異性且有效：siPOOLs在標準細胞系中將脫靶率降低5-25 倍，且在1-3 nM下實現(xiàn)基因敲低率≥70%。 

一致的表型：與siRNA相反，由序列非依賴性的siPOOL產生 的表型高度一致。 

確保經過檢驗：通過RT-qPCR進行siPOOL驗證，如果在 轉染下敲低率低于70%，則有可能重新設計。 

使用注釋進行定制設計：專業(yè)的設計，確保優(yōu)化熱力學特性且避免旁系 同源基因。 

HPLC純化且無毒：所有siPOOL均經過HPLC純化，可降低污染物 和副作用的風險。

關鍵問題：siRNAs的脫靶效應

科學家們一直在使用RNA干擾（RNAi）作為研究基因功能的快速有效的工具。然而，短干擾RNA（siRNA）的脫靶效應和可變性能仍然是一個令人頭疼的缺點，在驗證工作中消耗了寶貴的時間和資源。

siRNA通常與靶RNA轉錄本互補結合，通過RNAi機制指導其降解。脫靶效應主要是由siRNA模擬內源性 基因調節(jié)因子microRNA（miRNA）引起的。由于miRNA只需要6個堿基種子匹配到3'非翻譯區(qū)（UTR）即可觸 發(fā)轉錄本下調，因此siRNA在通過這種機制處理時可以改變許多意外靶標的表達。

siPOOLs如何提高特異性

單個siRNA或含有3-4個siRNA的低復雜度siRNA庫，經常擊中多個脫靶基因并表現(xiàn)出易變的靶基因敲低。 siPOOL是高度復雜且確定的30個siRNA池，每個siRNA以皮摩爾工作濃度存在。因此， · 稀釋了每個siRNA的脫靶特征，提高了靶向特異性。 · 確保了靶基因的協(xié)同敲低，產生更穩(wěn)健、更可靠的結果。

使用siPOOL具有更高的特異性

特異性好的試劑應僅影響其靶標基因。 

HeLa細胞中微陣列的表達譜分析顯示，單個siRNA可以誘導許多脫靶基因（紅點），而針對同一靶基因（綠 點）并包含非特異性siRNA的siPOOL大大降低了脫靶效應。

通過siPOOL實現(xiàn)更好的重現(xiàn)性和有效的敲低效率

普通siRNA的敲低效率差異很大。 

與單個siRNA相比，針對同一基因的siPOOLs具有相似的敲低效率，表明具有更高的穩(wěn)健性和可重復性（左 圖和中圖：靶RNA水平的實時定量PCR測量的相關圖）。 siPOOLs也表現(xiàn)出有效的基因敲低。在常用的細胞系中，許多基因通常在1 nM濃度下實現(xiàn)75-98%的基 因敲低效率（右圖）。

您可以信賴的表型

如果RNAi效果可靠且特異，靶向同一基因的兩種試劑應產生相似的表型。 

每個基因分別使用兩個siPOOL池，一個來自我們的人類激酶siPOOL文庫，一個是市售的每個基因含3條 siRNAs的文庫，科學家在A549細胞中篩選了36個基因并檢測了細胞活力。僅siPOOLs產生了較好的一致性表型。

siTOOLs生物技術始終致力于幫助學術研究人員、科學家、制藥/生物技術公司和RNAi篩選人員。目前，已幫助許多論文 發(fā)表在《自然》、《細胞》、《自然醫(yī)學》、《科學報告》等頂級期刊上。期待與您的合作，我們將為您的RNA研究提供量身定制 的分子工具。

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