zeta life转染造血干细胞(HPCs)共转染2种质粒DNA(IF11.556)论文分享

与t细胞技术有关的表面抗体(life转染造血干细胞)(1)

zeta life转染造血干细胞(HPCs)共转染2种质粒DNA(IF11.556)论文分享编辑

一、 转染造血干细胞(HPCs)或造血祖细胞-共转染两种质粒DNA文章概述:

检测特异性蛋白Sp1对FcγRIIB的影响在造血干细胞(HPCs )中表达,FcγRIIB 启动子区域为(-1500 ~ 1从转录起始位点),合成并亚克隆到pGL3载体, Sp1结合位点 5'-GGGGCGGGGC突变为 5'-AAGGCAAGGC,含有 Sp1过表达或含有模拟物。

使用美国Zeta Life公司Advanced DNA RNA转染试剂(#AD600025,Zeta Life, USA)对造血干细胞(HPCs )共转染 1 μg 报告载体和 20 ng pSV-Renilla 表达载体(含、模拟物和 Sp1),转染的细胞是孵育 48 小时后收细胞并检测。

文章作者:重庆大学附属肿瘤医院肿瘤内科IF11.556

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

二、转染造血干细胞(HPCs)

与t细胞技术有关的表面抗体(life转染造血干细胞)(2)

zeta life转染造血干细胞(HPCs)共转染2种质粒DNA(IF11.556)论文分享 编辑

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.

、美国Zeta Life 公司与美国加利福尼亚大学旧金山校区联合开发用于哺乳动物细胞、活体动物转染的 Advanced DNA RNA 第三代多肽小分子转染试剂,此技术成为新的全球蛋白功能、免疫细胞及干细胞治疗、研发及生产的主要关键技术之一。

、文献中所用转染试剂产品信息

产品名称:Advanced DNA RNA 转染试剂

货号:AD600025、AD600050、AD600075、AD600150

规格:0.25ML、0.5ML、0.75ML、1.5ML

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