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The cold tolerance mechanisms of insect have been studied extensively on the model species Drosophila and a few other species at the transcriptional level. However studies on insects that inherit strong cold tolerance are limited. Cold hardy Tenebrionid beetle Microdera punctipennis is endemic to Gurbantonggut Desert, northwest of China. However, its genomic information is lacking. To investigate the overwintering mechanisms of M. punctipennis adult, RNA-seq was performed on the winter adults and the control adults that were kept in laboratory at 30 degrees C. A total of 175,247 unigenes were acquired with an average length of 645 bp. By using DESeq package, we identified 3367 unigenes that were up-regulated and 7988 down-regulated in the winter adults compared with the controls. To further our understanding of these differentially expressed genes (DEGs), Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. Pathway analysis showed that the "ECM-receptor interaction", "PI3K-Akt signaling pathway", "Estrogen signaling pathway", "Tight junction", and "Regulation of actin cytoskeleton", etc. might play important roles in M. punctipennis overwintering. The DEGs results from the RNA-Seq were confirmed partially by qRT-PCR for 13 DEGs, which showed high consistence with a Pearson’s correlation coefficient of 0.851. Overall, the sequence data will provide basic information for subsequent bioinformatical analysis and mining of the genes responsible for cold tolerance in M. punctipennis, as well as for understanding the molecular mechanisms of desert beetle overwintering. (C) 2017 Elsevier Inc. All rights reserved.