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)

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。