Project description:Transcription profiling by array of Arabidopsis MKK9DD (constitutively active MKK9 kinase mutant) overexpressing seedlings and Pi-starved wild type seedlings to identify the same regulated genes
Project description:Transcript accumulation was measured using the Affymetrix Arabidopsis ATH1 Genome Array [ATH1-121501] to document changes in response to the MADS-domain transcription factor AGAMOUS-Like 15 during somatic embryogenesis. A somatic embryo system was used where mature seed is allowed to complete germination in liquid MS media containing 2,4-D and seedlings produce somatic embryos from the shoot apical meristem (SAM) region. The frequency with which these embryos are produced directly correlates with AGL15 accumulation.
Project description:Transcript profiling analysis of AtFBP7 mutant seedlings compared to wild type using Arabidopsis ATH1 GeneChip array. Keywords: 5 day old light grown seedlings, wild type and mutant
Project description:The Arabidopsis thaliana Myb transcription factor, FE, acts as a key regulator of phase transition. In order to identify potential target genes of FE protein, we performed microarray experiments. Using fe-1 and transgenic plants overexpressing GR-tagged FE (35S::FE-GR), we compared transcriptional profiling of WT (L.er) vs fe-1 and Dex-treated 35S::FE-GR vs Mock-treated 35S::FE-GR. Transcriptional profiling of A. thaliana comparing WT (L.er) with the fe-1 mutant
Project description:Somatic cells can be reprogrammed to induced pluripotent stem cells (iPSCs) by overexpressing OCT4, SOX2, KLF4 and MYC. Although this cell fate transition is a slow and inefficient process, inhibiting several epigenetic barriers has shown to facilitate it. A previously employed epigenetic focused CRISPR-Cas9-mediated knockout screen during reprogramming uncovered novel human somatic cell reprogramming barriers. Here, I showed that USP22, target for gRNAs that were enriched in iPSCs from the screen, is a barrier for reprogramming and its loss is compatible with iPSC generation. Overexpressing wild-type or deubiquitinase mutant (C185A) USP22 in knockout background could rescue the phenotype. Additionally, knocking out other SAGA deubiquitinase subunits, ENY2 and ATXN7L3 did not increase the reprogramming efficiency. Mechanistically, we showed that USP22 loss upregulates an important pluripotency-related transcription factor, SOX2, expression even as early as 3 days after OSKM expression. We hypothesized that USP22 downregulates pluripotency-related gene expression during reprogramming independent from its SAGA integration and deubiquitinase activity. To address this, I will repeat rescuing USP22 knockout phenotype in reprogramming by overexpressing mutant versions of USP22 including K129R (acetyl inhibited) and HHAA (deubiqutinase). Additionally, I will check the expression of pluripotency-related genes at early time points of reprogramming upon USP22 knockout by transcriptome profiling. Lastly, I will generate USP22 knockout iPSC lines to force them to differentiation to observe if they have any problem with pluripotency exit. This work will establish a mechanistic understanding for the role USP22 play on both human somatic cell reprogramming, pluripotency and differentiation.
Project description:Transcript profiling analysis of csn4-1 light grown mutant seedlings compared to wild type using Arabidopsis ATH1 GeneChip array Keywords: 7 day old light grown seedlings, wild type and mutant