Project description:CHQ5B primary human myoblasts obtained from Dr V. Mouly (Paris, France) have been described previously (Faulkner et al., 1999). Ankrd2 is upregulated on differentiation of myoblasts. The change in gene expression was studied in cells silenced for Ankrd2 (infected with AAV-shAnkrd2), control cells (infected with AAV-shLuc) and control cells uninfected.
Project description:NRAS and BRAF are the most commonly found genetic alterations in melanoma. It has been demonstrated that NRAS and BRAF oncogenic alterations, even affecting the same signaling pathway, represent two different clinical and biochemical entities, showing different signaling patterns and biological responses. Metabolic reprogramming is considered a novel target to control cancer; however, it is mostly unknown how the NRAS oncogene contributes to this cancer hallmark. We have showed that NRAS-mutated melanomas harbor specific metabolic alterations that render cells sensitive to RAS pathway inhibition upon metabolic stress. Gene expression analysis has confirmed different transcriptional profiles of NRAS- and BRAF- mutant cells both, under basal conditions and in response to glucose starvation. Moreover, analysis of glucose metabolism-related genes expression regulation revealed PFKFB2 as an important player linking glycolysis and RAS pathway activation.
Project description:The purpose of this study is to discern global expression programs embodying therapeutic targeting of CLN3 disease. Differential gene expression in Cln3Δex7/8 mouse brains was determined after intraperitoneal vehicle/Galactosylceramide (GalCer) injections, for 40 weeks, using GeneChip Mouse Genome 430 2.0 arrays. Differentially expressed genes among Cln3Δex7/8 mice and profiles modulated by GalCer treatment were functionally analyzed and topologically organized.
Project description:We are comparing the gene expression patterns from rats in which they are either double housed or socially isolated. Within these groups we performed the following manipulations:<br><br>AAV-GFP to the NAc shell<br><br>AAV-CREB to the NAc shell<br><br>chronic imipramine treatment<br><br>water control<br><br>
Project description:AAV is widely used for efficient delivery of DNA payloads. The extent to which the AAV capsid can be used to deliver a protein payload is unexplored. Here, we report engineered AAV capsids that directly package proteins – Protein Carrier AAV (pcAAV). Nanobodies inserted into the interior of the capsid mediate packaging of a cognate protein, including Green Fluorescent Protein (GFP), Streptococcus pyogenes Cas9, Cre recombinase, and the engineered peroxidase APEX2. We show that protein packaging efficiency is affected by the nanobody insertion position, the capsid protein isoform into which the nanobody is inserted, and the subcellular localization of the packaged protein during recombinant AAV capsid production; each of these factors can be rationally engineered to optimize protein packaging efficiency. We demonstrate that protein packaged within pcAAV retain their enzymatic activity and that pcAAV can bind and enter the cell to deliver the protein payload. Establishing pcAAV as a protein delivery platform expands the utility of AAV as a therapeutic and research tool.
Project description:We used AAV as a vector to deliver hGRβ to C57BL/6 mouse liver. We collected liver samples for microarray analysis to investigate any phenotype as well as the underlying specific signaling pathway. In particular, we would like to determine if and how hGRβ overexpression in liver affects mGRαâs gene transcription profile in C57BL/6 mice. Three replicates for each group: untreated WT liver, AAV-GFP treated liver, and AAV-hGRB treated liver.
Project description:We used AAV as a vector to deliver hGRβ to the liver of GR Liver Knockout mice. We collected liver samples for microarray analysis to investigate any phenotype as well as the underlying specific signaling pathway. In particular, we would like to determine if and how hGRβ overexpression in liver affects mGRαâs gene transcription profile in GR Liver Knockout mice. GR Liver Knockout mice were treated with PBS, AAV-GFP and AAV-hGRB, respectively. Each treatment group has four replicates.
Project description:The purpose of the experiment is to study the expression profiles variations upon the expression of the wild type isoform of LKB1 (STK11) in human lung carcinoma A459 cells. These cells are KRAS mutated and null for STK11. Cells were previously infected with the lentiviral construct pLenti-rtTA2-IRES-H2B-GFP doxycycline-inducible plasmid (obtained from S. Tenbaum, HG Palmer’s Lab Vall d´Hebron Institute of Oncology, VHIO) containing the human STK11wild type cDNA.
Project description:To study monocyte and macrophage activation in ANCA-associtated vasculitis (AAV), we performed bulk RNA sequencing of bead-selected monocytes and in vitro cultured monocyte-derived macrophages from AAV patients and healthy controls. Overview patients included for sequencing monocytes: - AAV active disease, n=4, MPO-AAV=4 - AAV remission, n=10, PR3-AAV=5, MPO-AAV=5 - Healthy controls, n=6 Overview patients included for sequencing monocyte-derived macrophages: - AAV active, n=1, PR3-AAV=1 - AAV remission, n=3, PR3-AAV=3 - Healthy controls, n=3