Project description:Pv11 is an insect cell line established from Polypedilum vanderplanki with ability to survive complete desiccation if they were treated with trehalose. Previous studies showed contribution of HSF1 gene in this adaptation. To determine HSF1 regulatory network in Pv11 cells we prepared RNAseq libraries for wild type Pv11 (WT), HSF1 knoked out (Hsf1-/-), HSf1 resqued (Hsf1-/-; Hsf1-HaloTag/BlaR), and negative control for HSF1 resqued cells (Hsf1-/- HaloTag/BlaR) under normal conditions and under incubation with trehalose for 48 h. Comparison of mRNA profiles showed that HSF1 induced anhydrobiosis-related genes, especially LEA and TRX, but suppressed cellular process-related genes, for example transcription- and translation-related genes.
Project description:Genomic safe harbors (GSHs) are utilized as an ideal integration site for generating transgenic organisms and cells. Discovery of GSHs is one of the crucial factors for the advancement of basic and applied biology in the species. Such GSHs were discovered in Pv11 (Polypedilum vanderplanki) cell line, which can survive extreme desiccation. To identify the integration sites, high-molecular-weight genomic DNAs were extracted. The DNA libraries were prepared and sequenced with a Nanopore MinION sequencer. In the way to confirm that GSHs loci are localized in open chromatin regions we prepared ATAC-seq libraries, which were sequenced in Illumina HiSeq2500.
