Project description:We profiled the transcriptomic changes that occur in the Drosophila third antennal segment after silencing the second order olfactory neurons in the antennal lobe with Kir2.1 (EH1). A non-conducting variant of Kir2.1 (Kir2.1-nc or EH2) served as our control.
Project description:Purpose:Mouse BMDM is the universal cell type of studying innate immunity.This study was to analyze LPS induced innate immune response and the relationship between the inward rectifier potassium channel Kir2.1 and LPS induced innate immune respone Methods: WT and Kcnj2 KO BMDM cells were treated with or without LPS for 6 hours in the presence of kir2.1 inhibitor ML133 or not. Then mRNA profiles of these samles were generated by High-throughput sequencing analysis, in triplicate, using illumina HiSeq 2000. And the differential mRNA profiles were analyzed. Results: mRNA profiles of more than 20000 genes were analyzed, and differential expression profiles were compared . Conclusions: Our study represents the first detailed analysis of transcriptomes of WT and Kcnj2 KO mouse BMDM treated with LPS or kir2.1 inhibitor, with biologic replicates, generated by RNA-seq technology. This data was useful to analyze the relationship between Kir2.1 and LPS induced innate immune respone
Project description:Kir2.1, an inward rectifier potassium channel, plays an important role in controlling membrane potential specially in cardiomyocytes. Here we explore its interactome via proximity biotin labeling and affinity purification approach (BioID). First, BioID membrane controls were generated with randomly selected transmembrane domains from yeast proteins, demonstrating the identification of membrane-associated proteins more than GFP or NLS controls. Using this control and a mutation from Andersen-Tawil syndrome which causes Kir2.1 Golgi accumulation, we have identified the most comprehensive Kir2.1 interactome and also found clues to its spatial information in subcellular levels. The interactome is showing that Kir2.1 in plasma membrane is surrounded by desmosome, integrin, cadherin and dystrophin-glycoprotein complex complexes, and supported by MAGUK family scaffold proteins. On the other hand, Kir2.1 mutant BioID revealed the COPII-mediated delivery of Kir2.1 from ER to Golgi and lysosome-mediated degradation of Golgi-accumulated and/or retrograde Kir2.1. Validating the interactome with co-immunoprecipitation, confocal microscope, and patch clamp analysis, we concluded that Kir2.1 is located in the defined membrane environment and is actively regulated by endosomal sorting for lysosome degradation.
Project description:In order to characterize precisely the T-DNA insertion(s) in three reporter lines, we performed whole genome sequencing and searched for insertion sites and T-DNA copy numbers.
Project description:Tn insertion library was used for recipient for conjugative transfer of pESBL, F, and R388 plasmids. For both recipient and the resulting exconjugant libraries, Tn insertion sites were determined by illumina sequencing
Project description:Transposon insertion site sequencing (TIS) is a powerful method for associating genotype to phenotype. However, all TIS methods described to date use short nucleotide sequence reads which cannot uniquely determine the locations of transposon insertions within repeating genomic sequences where the repeat units are longer than the sequence read length. To overcome this limitation, we have developed a TIS method using Oxford Nanopore sequencing technology that generates and uses long nucleotide sequence reads; we have called this method LoRTIS (Long Read Transposon Insertion-site Sequencing). This experiment data contains sequence files generated using Nanopore and Illumina platforms. Biotin1308.fastq.gz and Biotin2508.fastq.gz are fastq files generated from nanopore technology. Rep1-Tn.fastq.gz and Rep1-Tn.fastq.gz are fastq files generated using Illumina platform. In this study, we have compared the efficiency of two methods in identification of transposon insertion sites.