Project description:Since radiotherapy is the essential treatment of prostate cancer, this experiment aimed to elucidate the transcriptomic response of prostate cancer cells to irradiation by either conventional photons or alternatively carbon ions, focusing on DNA damage, DNA repair and androgen receptor signaling.
Project description:p53 expression and transcriptional activity is repressed in human papillomavirus positive cells by the expression of viral E6 protein. E6 forms a complex with cellular E6AP protein which can lead to ubiquitination of p53, thus repressing its tumor-suppressive capabilities. In this experiment, the potential capability of a DARPin (Designed Ankyrin Repeat Protein) to reactivate p53 mediated pathways in HPV positive HeLA cells is investigated and compared to Nutlin 3a activation of p53 in HPV negative U2OS osteosarcoma cells.
Project description:Peripheral blood monocytes were differentiated into macrophages, starved for 24h hours, and treated for 3h with LPA 18:0, 20:4 or a mix of LPAs similar to the composition in ascites (1 µM 16:0 + 0.25 µm 18:0 + 0.5 µM 18:1 + 1.625 µM 18:2 + 1.625 µM 20:4). RNA expression was measured via QuantSeq.
Project description:To compare the impact of several TP53 mutant variants in an isogenic setting, different TP53 mutations were introduced in HCT116 colorectal carcinoma cells. This parental cell line is wild-type for TP53 and shows a prototypical p53 response. To ensure unambigous genotype-phenotype correlations, the cell were haploidized prior to CRISPR-editing by introducing inactivating deletions of intronic splicing into one of the two TP53 alleles, leaving only one functional copy of TP53. The remaining TP53 allele was altered by inserting a LoxP-flanked transcriptional stop cassette (Lox-Stop-Lox, LSL) into intron 4, which allowed reversible silencing of TP53 expression. The LSL cassette was then specifically targeted with CRISPR/Cas9 to introduce a variety of different mutant p53 alleles. The competency of the mutated p53 allele to induce a p53 response upon activation using Nutlin-3a was then assessed in an RNAseq experiment.
Project description:QuantSeq of immortalised primary human foetal foreskin fibroblasts (HFFF-TERTs) infected with Sendai Virus (SeV) or mock infected. Experiments were performed in two rounds. In the first round cells were depleted of both CRTC2 and CRTC3 using siRNAs, or treated with a control siRNA. In the second round of experiments HFFF-TERT cells had either CRTC2, CRTC3 or both CRTC2&CRTC3 knocked out using a CRISPR-Cas9 system. Knock-out cells were complemented with either full length CRTC2, full length CRTC3, a version of CRTC2 lacking the N-terminal 50 amino acids (ΔN50), a version of CRTC3 lacking the N-terminal 50 amino acids (ΔN50), both full length CRTC2 and CRTC3, or a vector only control.
Project description:The aim of this experiment was to assess the on- and off-target effects of MAPT-AS1 expression, and whether mutations/deletions to MAPT-AS1 alter these effects. SHSY5Y cells stably expressing variants of MAPT-AS1 were analyzed by Riboseq and Quantseq.