zeta life轉染造血幹細胞(HPCs)共轉染2種質粒DNA（IF11.556）論文分享

zeta life轉染造血幹細胞(HPCs)共轉染2種質粒DNA（IF11.556）論文分享編輯

一、 轉染造血幹細胞(HPCs)或造血祖細胞-共轉染兩種質粒DNA文章概述：

檢測特異性蛋白Sp1對FcγRIIB的影響在造血幹細胞（HPCs ）中表達，FcγRIIB 啟動子區域為(-1500 ~ 1從轉錄起始位點)，合成并亞克隆到pGL3載體, Sp1結合位點 5'-GGGGCGGGGC突變為 5'-AAGGCAAGGC，含有 Sp1過表達或含有模拟物。

使用美國Zeta Life公司Advanced DNA RNA轉染試劑（#AD600025，Zeta Life, USA）對造血幹細胞（HPCs ）共轉染 1 μg 報告載體和 20 ng pSV-Renilla 表達載體（含、模拟物和 Sp1），轉染的細胞是孵育 48 小時後收細胞并檢測。

文章作者：重慶大學附屬腫瘤醫院腫瘤内科IF11.556

标題：FcγRIIB potentiates differentiation of myeloid-derived suppressor cells to mediate tumor immunoescape

二、轉染造血幹細胞(HPCs)

zeta life轉染造血幹細胞(HPCs)共轉染2種質粒DNA（IF11.556）論文分享 編輯

FcγRIIB promoter luciferase reporter assay

To measure the effect of Sp1 on FcγRIIBexpression in HPCs, the FcγRIIB promoter region

(-1500 ~ 1 from the transcription starting site) wassynthesized by GenScript co., LTD (Nanjing, China)and subcloned into pGL3-basic vector (Promega,Madison, WI, USA), Sp1 binding site 5’-GGGGCGGGGC was mutated to 5’-AAGGCAAGGC.Lentivectors containing Sp1 overexpression or a lentivector containing mock were obtained from Genechem (Shanghai, China). The HPCs were transfected with 1 μg of reporter vector and 20 ng of pSV-Renilla expression vector (mock and Sp1) using Advanced DNA RNA Transfection Reagent (Cat No.AD600025, Zeta-life). Transfected cells were incubated for 48 h, Luciferase and Renilla activitie were measured using the dual-luciferase reporter system kit (Cat No. E1910, Promega). The transfection was performed according to the manufacturer' protocol.

三、美國Zeta Life 公司與美國加利福尼亞大學舊金山校區聯合開發用于哺乳動物細胞、活體動物轉染的 Advanced DNA RNA 第三代多肽小分子轉染試劑，此技術成為新的全球蛋白功能、免疫細胞及幹細胞治療、研發及生産的主要關鍵技術之一。

四、文獻中所用轉染試劑産品信息

産品名稱：Advanced DNA RNA 轉染試劑

貨号：AD600025、AD600050、AD600075、AD600150

規格：0.25ML、0.5ML、0.75ML、1.5ML