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技術文章
zeta轉染造血干細胞(HPCs)或造血祖細胞-共轉染2種質粒DNA
閱讀：618 發(fā)布時間：2022-4-13
一、 轉染造血干細胞(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的影響在造血干細胞（HPCs ）中表達，F(xiàn)cγRIIB 啟動子區(qū)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?yōu)?5'-AAGGCAAGGC，含有 Sp1過表達或含有模擬物。
使用美國Zeta Life公司Advanced DNA RNA轉染試劑（#AD600025，Zeta Life, USA）對造血干細胞（HPCs ）共轉染 1 μg 報告載體和 20 ng pSV-Renilla 表達載體（含、模擬物和 Sp1），轉染的細胞是孵育 48 小時后收細胞并檢測。
文章作者：重慶大學附屬腫瘤醫(yī)院腫瘤內科IF11.556
標題：FcγRIIB potentiates differentiation of myeloid-derived suppressor cells to mediate tumor immunoescape
二、轉染造血干細胞(HPCs)
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(qū)聯(lián)合開發(fā)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，此技術成為新的蛋白功能、免疫細胞及干細胞治療、研發(fā)及生產的主要關鍵技術之一。
四、產品信息
