mRNA expression in Zfp36L2 knockout E14.5 fetal liver

ZFP36L2, zinc finger protein 36, C3H type-like 2 (also known as Brf2, Erf2, Tis11D) is a member of the tristetraprolin (TTP; Zfp36) family of tandem CCCH zinc finger proteins that can bind to AU-rich elements (AREs) in the 3'-untranslated region of mRNAs, leading to their deadenylation and subsequent degradation. We have generated Zfp36l2 knockout mice. Knockout mice were born at the expected Mendelian frequency, but within several weeks of birth they died rather suddenly with pallor and frequent intestinal hemorrhage. These mice exhibited pancytopenia, decreased hematopoietic progenitor cells from fetal liver and yolk sac, and ineffective hematopoietic stem cells. Since ZFP26L2 is likely to function as an ARE-containing mRNA destabilizing protein, we were interested in identifying any abnormally stabilized transcripts in fetal livers from the Zfp36l2 knockout mice whose protein product may directly or indirectly affect hematopoietic stem cell function. Total RNA was isolated from individual embryonic day 14.5 fetal livers. Five wild type and five knockout fetal livers were used to generate 5 wild type replicates and 5 knockout replicates. RNA expression from knockout fetal livers was compared to wild type fetal livers.ZFP36L2（锌指蛋白36 C3H型样蛋白2，Zinc finger protein 36, C3H type-like 2，又名Brf2、Erf2、Tis11D）属于三联体促凋亡蛋白（tristetraprolin，TTP；Zfp36）家族，该家族为串联CCCH型锌指蛋白，可结合mRNA 3'非翻译区的AU富集元件（AU-rich elements，AREs），介导mRNA的脱腺苷酸化及后续降解。本研究构建了Zfp36l2基因敲除小鼠。敲除小鼠以预期的孟德尔遗传频率出生，但在出生后数周内突然死亡，表现为面色苍白并伴随频发肠道出血。该类小鼠呈现全血细胞减少症，胎肝与卵黄囊中的造血祖细胞数量减少，且造血干细胞功能无效。鉴于ZFP26L2大概率作为含ARE的mRNA不稳定化蛋白发挥功能，我们旨在鉴定Zfp36l2基因敲除小鼠胎肝中异常稳定的转录本，此类转录本的编码蛋白可能直接或间接影响造血干细胞功能。研究人员从单只胚胎发育第14.5天的胎肝中分离总RNA，选取5份野生型胎肝与5份敲除型胎肝，分别构建5个野生型重复样本与5个敲除型重复样本，将敲除型胎肝的RNA表达水平与野生型胎肝进行比较。