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轉(zhuǎn)染造血干細(xì)胞(HPCs)或造血祖細(xì)胞-共轉(zhuǎn)染2種質(zhì)粒DNA文章概述
閱讀：962 發(fā)布時間：2022-5-19
一、 轉(zhuǎn)染造血干細(xì)胞(HPCs)或造血祖細(xì)胞-共轉(zhuǎn)染兩種質(zhì)粒DNA文章概述：
檢測特異性蛋白Sp1對FcγRIIB的影響在造血干細(xì)胞（HPCs ）中表達(dá)，F(xiàn)cγRIIB 啟動子區(qū)域為(-1500 ~ +1從轉(zhuǎn)錄起始位點)，合成并亞克隆到pGL3載體, Sp1結(jié)合位點 5'-GGGGCGGGGC突變?yōu)?5'-AAGGCAAGGC，含有 Sp1過表達(dá)或含有模擬物。
使用美國Zeta Life公司Advanced DNA RNA轉(zhuǎn)染試劑（#AD600025，Zeta Life, USA）對造血干細(xì)胞（HPCs ）共轉(zhuǎn)染 1 μg 報告載體和 20 ng pSV-Renilla 表達(dá)載體（含、模擬物和 Sp1），轉(zhuǎn)染的細(xì)胞是孵育 48 小時后收細(xì)胞并檢測。
文章作者：重慶大學(xué)附屬腫瘤醫(yī)院腫瘤內(nèi)科IF11.556
標(biāo)題：FcγRIIB potentiates differentiation of myeloid-derived suppressor cells to mediate tumor immunoescape
二、轉(zhuǎn)染造血干細(xì)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(xué)舊金山校區(qū)聯(lián)合開發(fā)用于哺乳動物細(xì)胞、活體動物轉(zhuǎn)染的 Advanced DNA RNA 第三代多肽小分子轉(zhuǎn)染試劑，此技術(shù)成為新的蛋白功能、免疫細(xì)胞及干細(xì)胞治療、研發(fā)及生產(chǎn)的主要關(guān)鍵技術(shù)之一。
四、產(chǎn)品信息
一、 轉(zhuǎn)染造血干細(xì)胞(HPCs)或造血祖細(xì)胞-共轉(zhuǎn)染兩種質(zhì)粒DNA文章概述：
檢測特異性蛋白Sp1對FcγRIIB的影響在造血干細(xì)胞（HPCs ）中表達(dá)，F(xiàn)cγRIIB 啟動子區(qū)域為(-1500 ~ +1從轉(zhuǎn)錄起始位點)，合成并亞克隆到pGL3載體, Sp1結(jié)合位點 5'-GGGGCGGGGC突變?yōu)?5'-AAGGCAAGGC，含有 Sp1過表達(dá)或含有模擬物。
使用美國Zeta Life公司Advanced DNA RNA轉(zhuǎn)染試劑（#AD600025，Zeta Life, USA）對造血干細(xì)胞（HPCs ）共轉(zhuǎn)染 1 μg 報告載體和 20 ng pSV-Renilla 表達(dá)載體（含、模擬物和 Sp1），轉(zhuǎn)染的細(xì)胞是孵育 48 小時后收細(xì)胞并檢測。
文章作者：重慶大學(xué)附屬腫瘤醫(yī)院腫瘤內(nèi)科IF11.556
標(biāo)題：FcγRIIB potentiates differentiation of myeloid-derived suppressor cells to mediate tumor immunoescape
二、轉(zhuǎn)染造血干細(xì)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(xué)舊金山校區(qū)聯(lián)合開發(fā)用于哺乳動物細(xì)胞、活體動物轉(zhuǎn)染的 Advanced DNA RNA 第三代多肽小分子轉(zhuǎn)染試劑，此技術(shù)成為新的蛋白功能、免疫細(xì)胞及干細(xì)胞治療、研發(fā)及生產(chǎn)的主要關(guān)鍵技術(shù)之一。
四、產(chǎn)品信息