Project description:ABA regulates in plants a wide range of developmental events, mediates responses to environmental stress and is necessary to proceed through seed maturation and to acquire desiccation tolerance and dormancy. Immuno-modulation is a suitable means to study ABA functions during seed maturation. Anti-ABA single chain antibody was expressed in pea seed driving LeB4-promoter (Saalbach et al., High-level expression of a single chain Fv fragment (scFv) antibody in transgenic pea seeds J. Plant Physiol. 2001 158: 529-533), which produced only a weak phenotype with slightly decreased seed weight, globulin/albumin and total nitrogen content (aABA line 16 cultivar Erbi). In another approach with a stronger, improved USP-promoter used to express the anti-ABA antibody in pea seeds a different phenotype emerged (aABA line 7, cultivar Eifel). In this line individual seed weight increased by 20 to 30% together with higher globulin and albumin content. To dissect the aABA phenotype at the molecular level, a search for genes with differential expression patterns in transgenic plant versus wild type seeds has been performed using 6k-oligo microarray analysis. cDNA probes were prepared from RNA isolated from embryo of developing seeds of wild type (12, 18, and 22 DAP) and transgenic aABA plants (12, 18, and 22 DAP), which correspond to the transition phase of seed development, and 6k-oligo microarray.
Project description:Alterations of the crosstalk between the nervous and immune systems may underlie the inflammatory and neurodegenerative changes characterizing Multiple Sclerosis (MS). Among the molecules that actively interconnect these two systems is the Nerve Growth Factor (NGF), involved in both regulating inflammation and in brain homeostasis. Additionally, NGF is known to modulate oligodendrogenesis and reduce myelination, further suggesting an involvement of NGF in MS ethiology. In this study, we find that NGF regulates oligodendrogenesis by modulating the levels of miR-219a-5p, a well-known positive regulator of oligodendrocyte (OL) differentiation. We utilized a transgenic NGF-deprivation mouse model, AD11, in which postnatal expression of an anti-NGF antibody leads to NGF neutralization and progressive neurodegeneration. Notably, we find that these mice also display increased myelination. A microRNA profiling of AD11 brain samples (hippocampus) by Agilent microarray analysis revealed that NGF deprivation leads to an increase of miR-219a-5p levels and a downregulation of its predicted targets, compared to AD11-VH control mice. The AD11-VH controls are transgenic mice where only the heavy chain of the transgenic antibody is expressed, while in AD11 mice express the whole functional antibody (heavy + light chain)
Project description:LAIR1 is an inhibitory immune cell receptor. We used single cell RNA sequencing (scRNA-seq) to analyze the immune cell distribution in the implanted B16 tumor tissues of Vav-cre human LAIR1 transgenic and LAIR1-/- C57bl/6j mice after control or anti-LAIR1 blocking antibody treatment.
Project description:Cell lines were generated by transfecting NS0 myeloma cells with a single vector which contained the genes for the heavy and light chain of the antibody anti-CD38, and also the selectable marker glutamine synthetase. After two rounds of limiting dilution cloning, long-term continuous culture was performed on two cell lines (4H8 and 4G3) to assess the stability of recombinant antibody production during periods of extended culture. Microarray analysis was performed on RNA samples extracted during log phase of growth at the start of long-term culture and after approximately one month.
Project description:We acquied the RNA polymerase II binding profiles in ama-1 transgenic worms. The large subuint of POL II, AMA-1, was C-terminally tagged with GFP, the binding regions were determined using chromatin immunoprecipitation with anti-GFP and anti-polymerase II antibodies followed by illumina high-throughput sequencing (ChIP-seq). For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf RNA polymerase II binding profiles at L4/young adult stage were acquied with anti-native protein antibody (8WG16) and anti-epitope (GFP) antibody