Project description:Taf4b-deficient male mice are initially sub-fertile and become infertile due to a depletion of the spermatogonial stem cell (SSC) reserve. During embryonic time points, significantly reduced numbers of germ cells have been observed in the Taf4b-deficient male gonad and previous research has shown that Taf4b mRNA expression peaks at E15.5. Therefore, we hypothesized that TAF4b, as part of TFIID, regulates cell cycle and SSC gene programs. However, the direct targets of TAF4b have not been thoroughly explored. Therefore, we performed Cleavage Under Targets and Release Using Nuclease (CUT&RUN) in E16.5 mouse prospermatogonia.
Project description:Research has shown that Taf4b-deficient female mice display excessive perinatal germ cell death, delayed germ cell cyst breakdown, and increased chromosome asynapsis. Therefore, we hypothesized that TAF4b, as part of TFIID, regulates oogenesis and meiotic gene programs. However, the direct targets of TAF4b have not been thoroughly explored. Therefore, we performed Cleavage Under Targets and Release Using Nuclease (CUT&RUN) in E16.5 mouse oocytes.
Project description:Taf4b-deficient male mice are initially sub-fertile and become infertile due to a depletion of the spermatogonial stem cell (SSC) reserve. During embryonic time points, significantly reduced numbers of germ cells have been observed in the Taf4b-deficient male gonad and previous research has shown that Taf4b mRNA expression peaks at E15.5. Therefore, we hypothesized that TAF4b, as part of TFIID, regulates cell cycle and SSC gene programs. We performed RNA-seq to examine gene expression changes in E14.5 and E16.5 male Taf4b-wildtype, Taf4b-heterozygous, and Taf4b-deficient germ cells
Project description:Research has shown that Taf4b-deficient female mice display excessive perinatal germ cell death, delayed germ cell cyst breakdown, and increased chromosome asynapsis. Therefore, we hypothesized that TAF4b, as part of TFIID, regulates oogenesis and meiotic gene programs. However, the transcriptomic effects of Taf4b-deficiency and how this may lead to the infertility we observe in mice has not yet been studied. Therefore, we performed RNA-seq to examine gene expression changes in E16.5 female Taf4b-heterozygous and Taf4b-deficient germ cells
Project description:Research has shown that Taf4b-deficient female mice display excessive perinatal germ cell death, delayed germ cell cyst breakdown, and increased chromosome asynapsis. Therefore, we hypothesized that TAF4b, as part of TFIID, regulates oogenesis and meiotic gene programs. However, the transcriptomic effects of Taf4b-deficiency and how this may lead to the infertility we observe in mice has not yet been studied. Therefore, we performed RNA-seq to examine gene expression changes in E14.5 female Taf4b-wildtype, Taf4b-heterozygous, and Taf4b-deficient germ cells
Project description:Expression analysis was performed with two TDNA insertion mutants of taf4b i.e; taf4bprm (TDNA insertion in promoter region) and taf4bint (TDNA insertion in intronic region), Taf4b overexpression lines, taf4bprmcpr5 double mutant lines (Double mutant was generated by crossing taf4bprm with cpr5) and Col-0 in normal condition as well as with taf4bprm mutant and Col-0 infected with fungi AB (Alternaria brassicicola) and bacteria ES4 (Pseudomonas syringae pv.maculicola ES4326 ) in different perspectives. Affymatrix expression analysis was executed to provide mechanistic details of regulation of genes by Taf4b in plants.
Project description:MBD2 genome-binding landscape was assessed by CUT&RUN-sequencing in differentiating C2C12 cells. H3K4Me3 CUT&RUN-sequencing was performed as a positive control. Negative control experiment was also performed using a rabbit isotype control monoclonal IgG.