In situ depletion of Nuak1 in murine colon tumours

The aim of the project is to determine the impact of NUAK1 depletion on the transcriptome of in situ colon tumours. Tumours were initiated in Villin-CRE-ER;APC-floxed;DI-shNUAK1 mice, followed by treatment for 1 week with dextran sodium sulphate. 68 days after tumour initiation, NUAK1 shRNAs were induced by doxycycline administration (via gavage), or left un-induced. 6 tumours were harvested from NUAK1 shRNA expressing mice and 6 from controls. RNA was isolated using Qiagen RNEasy and analyzed by Illumina paired-end RNA-SEQ

本研究旨在探究NUAK1敲降对原位结肠肿瘤转录组（transcriptome）的影响。实验在Villin-CRE-ER;APC-floxed;DI-shNUAK1小鼠中诱导结肠肿瘤发生，随后给予葡聚糖硫酸钠（dextran sodium sulphate）处理1周。肿瘤诱导发生68天后，通过灌胃给药多西环素（doxycycline）诱导NUAK1短发夹RNA（short hairpin RNA，shRNA）的表达，对照组则不进行诱导处理。分别从NUAK1 shRNA表达组与对照组小鼠中各收获6枚肿瘤组织。使用凯杰（Qiagen）RNeasy试剂盒提取总RNA，并通过Illumina双端RNA测序（RNA-SEQ）完成转录组分析。