Project description:Human prostate cancer LNCaP cells infected with Lentivirus carried huamn BRCA2 gRNA alone or in combination with lentivirus carrying human RB1 shRNA. Infected cell were selcted by drug selection and the knockdown were confirm by westernblot or qPCR
Project description:The goal of this study was to perform transcriptomics on wildtype, PTEN single knockout (SKO) and PTEN;Rb1 double knockout (DKO) mouse prostate organoids. We isolated basal cells from PTEN floxed and PTEN;Rb1 floxed mouse prostates and infected with either RFP control or Cre recombinase to establish wildtype, SKO, and DKO mouse prostate organoids.
Project description:Microarray-based expression profiling of BRCA2 knockout and isogenic wild type HCT116 human colorectal cancer cells One way ANOVA, single factor comparison of wild type and BRCA2 knockout cells, three CEL files for wild type, three CEL files for BRCA2 knockout
Project description:A549 cells were co-transfected with CRISPR/Cas9 (containing sgRNA and GFP) and HDR donor plasmids (containing RFP and puromycin). The genetic editing cells were selected by puromycin. And then, we performed RNA sequencing for transcription profiling of A549 RB1 RB1 wild-type/knockout cell lines. The raw data were processed with Tophat and cuffdiff for gene expression analysis.
Project description:The goal of this study was to determine how pharmacological inhibition of the mitochondrial pyruvate carrier (MPC) alters lineage identity in transformed prostate organoids. We isolated basal cells from PTEN,Rb1 floxed mouse prostates and infected with Cre recombinase to establish PTEN, Rb1 double knockout (DKO) mouse prostate organoids. We then treated the DKO organoids with vehicle or 10uM UK5099 for five days.
Project description:We found a high frequency of heterozygous Fanconi-BRCA pathway mutations in pediatric T-ALL. BRCA2 was the most commonly mutated gene. We transduced Cas9-expressing Jurkat cells, which lacked an identifiable BRCA2 mutation, with an integration-defective lentiviral guide RNA expression construct targeting exon 11 of BRCA2 (NM_000059). Single-cell cloning and sequencing analysis revealed two distinct clones harboring monoallelic BRCA2 frameshift mutations, termed clones W4 and W5. Each of these clones was subjected to RNA sequencing analysis.