RNA-expression profiling of LMP1 expressing mouse B cells before and after transformation in vivo. RNA-expression profiling of LMP1 expressing mouse B cells before and after transformation in vivo

Non-transformed LMP1 expressing mouse B cells or mouse LMP1 lymphomas were analysed by RNA-sequencing Overall design: 14 Rag2KOcgKO were reconstituted with fetal liver hematopoietic stem and progenitor cells (HSPC) from Cd3eKO Cd19-Cre R26LMP1stopf mice. From 3 mice LMP1 expressing non-transformed B cells were isolated day 18-20 post reconstitution. 11 mice developed LMP1-driven B-cell lymphomas. Thse lymphomas were transplanted into Rag2KOcgKO or Rag2KO mice. Arising secondary lymphomas were FACSpurified (CD19+huCD2+). RNA of sorted lymphoma samples was analysed by RNA sequencing.

本研究通过RNA测序（RNA-sequencing）分析了未转化的表达潜伏膜蛋白1（LMP1）的小鼠B细胞以及小鼠LMP1淋巴瘤。
总体实验设计：将14只Rag2敲除且γc敲除（Rag2KOcgKO）小鼠以来自Cd3ε敲除（Cd3eKO）、Cd19-Cre介导R26LMP1stopf小鼠的胎肝造血干细胞与祖细胞（hematopoietic stem and progenitor cells, HSPC）进行造血重建。其中3只小鼠在造血重建后第18~20天分离得到表达LMP1的未转化B细胞；剩余11只小鼠发生LMP1驱动的B细胞淋巴瘤。将这些淋巴瘤移植至Rag2KOcgKO或Rag2KO小鼠体内，获取的继发性淋巴瘤经荧光激活细胞分选（fluorescence-activated cell sorting, FACS）纯化得到CD19+huCD2+细胞群。随后对分选得到的淋巴瘤样本的RNA进行RNA测序分析。