Project description:We generated human induced pluripotent stem cell (iPSC) lines with a GFP reporter inserted in the endogenous NKX6.1 locus. Characterisation of the reporter lines demonstrated faithful GFP labelling of NKX6.1 expression during pancreas and motor neuron differentiation. We performed three independent in vitro differentiations towards the pancreatic endocrine lineage. We FACS-purified GFP positive and negative cells from stage 7 cultures, and generated Smart-Seq2 RNA-sequencing libraries for the pre-sorted cells, as well as the two GFP-sorted cell populations. Gene expression profiling by RNA-sequencing reveals that the NKX6.1-positive population closely resembles mature human beta cells and the functional evaluation of purified populations shows that the glucose-responsive beta-like cells are enriched within the NKX6.1-positive population. These reporter lines provide a valuable resource to the scientific community for the derivation of functional relevant pancreas and neuronal cell subtypes.
Project description:We generated human induced pluripotent stem cell (iPSC) lines with a GFP reporter inserted in the endogenous NKX6.1 locus. Characterisation of the reporter lines demonstrated faithful GFP labelling of NKX6.1 expression during pancreas and motor neuron differentiation. We performed three independent in vitro differentiations towards the pancreatic endocrine lineage. We FACS-purified GFP positive and negative cells from stage 7 cultures, and generated Smart-Seq2 RNA-sequencing libraries for the pre-sorted cells, as well as the two GFP-sorted cell populations. Gene expression profiling by RNA-sequencing reveals that the NKX6.1-positive population closely resembles mature human beta cells and the functional evaluation of purified populations shows that the glucose-responsive beta-like cells are enriched within the NKX6.1-positive population. These reporter lines provide a valuable resource to the scientific community for the derivation of functional relevant pancreas and neuronal cell subtypes.
Project description:Interventions: Nucleic acid from tumor tissues and serum SNPs
Primary outcome(s): 1. Tissue biomarkers including mutation, gene expression, and DNA methylation that correlate with efficacy from trifluridine/tipiracil hydrochloride therapy 2. Serum biomarkers including mutation, gene expression, and DNA methylation that correlate with efficacy from trifluridine/tipiracil hydrochloride therapy 3. SNPs that correlate with toxicities from trifluridine/tipiracil hydrochloride therapy
Study Design: Single arm Non-randomized
Project description:Regulation of gene expression at the translational level is key to determining cell fate and function. An RNA-binding protein RNG140 (caprin2) plays a role in cell differentiation such as eye lens development and has been reported to function in translational regulation. To identify RNG140-associated proteins, we generated CHO cells stably expressing GFP and RNG140-GFP, and performed immunoprecipitation (IP) with GFP antibody from those cells followed by mass spectrometry. We identified about 400 proteins specifically co-immunoprecipitated with RNG140-GFP but not with GFP. Gene ontology (GO) enrichment analysis revealed that the eIF3 subunit proteins and small ribosomal subunit proteins were significantly enriched in the RNG140-associated complex.
Project description:Gene expression profiling of GFP-positive cancer stem-like cells(CSLC) derived from MDA-MB453 cells, compared with GFP-negative non-cancer stem cells (non-CSLC).