Project description:Perturbation of host physiology by intracellular Mycobacterium tuberculosis (H37Rv) can be reflected by changes in gene expression pattern of host genes. Total RNA was isolated from PMA differentiated uninfected cells or cells infected with H37Rv for 16, 48 or 90 hours and gene expression profile was obtained. There are in all 8 samples, two replcates of each 4. Two samples, namely 1D,1E (replicates of one) are control (PMA differentiated Thp-1 cells Uninfected controls). Cells were infected with H37Rv at an MOI of 1:10 and samples were collected at 16 hours, 48 hours and 90 hours post-infection.
Project description:①Background:Tuberculosis is mainly a respiratory tract infection caused by mycobacterium tuberculosis and one of the leading causes of death worldwide. According to the Global Tuberculosis Report in 2021, About a quarter of the world's population is infected with Mycobacterium tuberculosis and China is the second highest burden of TB. Although TB diagnosis and prevention techniques have become more mature, the number of TB cases is still increasing, mainly due to: the prevalence of drug-resistant tuberculosis bacteria, tuberculosis and HIV co-infection, long incubation time of mycobacterium tuberculosis difficult to early diagnosis and so on. Therefore, it is of great significance to study the pathogenesis of mycobacterium tuberculosis infection.②Method: THP-1 cells were treated with 50ng/ml PMA for 24 hours, so that THP-1 cell can be induced into macrophages. After that THP-1 macrophages were infected with mycobacterium tuberculosis H37Rv(MOI=1), which were collected and applied to RNA-sequencing. The constructed sequencing library was sequenced using an Illumina Novaseq 6000 system.
Project description:Transcriptional profiling of Mtb H37Rv infected into THP-1 macrophage cell line and treated with 100 µM vitamin C (vit C) for 96 hours and 144 hours, compared to gene expression profile of untreated bacteria post-infection.
Project description:In this study, RNA sequencing (Transcriptome sequencing) was employed to investigate the global transcriptome changes in the macrophages during the different strains of M.tb infection. THP-1 cells derived macrophages were exposed to the virulent M.tb strain H37Rv (Rv) or the avirulent M.tb strain H37Ra (Ra), and the M.tb BCG vaccine strain was used as a control. The cDNA libraries were prepared from M.tb infected macrophages and then sequenced.
Project description:Differentially regulated miRNA candidates in H37Rv infected THP-1 cells were analysed with respect to uninfected THP-1 reference samples. THP-1 cells are monocytes differentiated to macrophages after treatment with PMA for 48 hrs. Total RNA was isolated from infected THP-1 cells after 24 hrs of infection, cDNA was synthesized for TLDA real time PCR reaction using TaqMan MicroRNA Reverse Transcription kit and Megaplex Human Pool A and Pool B stem loop RT primers (version 3.0) as per manufacturer’s protocol. Further real time reaction was performed on QuantStudio 12K Flex Real-Time PCR System (Applied Biosystems) by using cDNA (without pre-amplification) on TLDA card A and card B (version 3.0).
Project description:To explore the regulatory network of noncoding RNAs after M.tb infection and the role of Rv1759c in the infection process, we collected samples of H37Rv- and H37Rv△1759c-infected macrophages and explored the full transcriptome expression profile. We constructed DE-lncRNA/DE-circRNA-DE-miRNA-DE-mRNA regulatory networks during H37Rv and H37Rv△1759c infection. In addition, we first discovered the close relationship between Rv1759c and chemokines during M.tb infection by comparing the transcription profiles of H37Rv and H37Rv△1759c and bioinformatics analysis. Here, our study comprehensively characterizes the ncRNA and mRNA profiles in macrophages infected with H37Rv and H37Rv△1759c, which provides support and new directions for in-depth exploration of ncRNA and PE/PPE family functions during the infection process.
Project description:Perturbation of host physiology by intracellular Mycobacterium tuberculosis (H37Rv) can be reflected by changes in gene expression pattern of host genes. Total RNA was isolated from PMA differentiated uninfected cells or cells infected with H37Rv for 16, 48 or 90 hours and gene expression profile was obtained.