Project description:Sustained spermatogenesis in adult males and recovery of fertility following germ cell depletion are dependent on undifferentiated spermatogonia with self-renewal potential. Self-renewal of undifferentiated spermatogonia is dependent on a complex network of transcriptional and post-transcriptional regulatory factors. DDX5 is known to act as a transcriptional co-regulator in other systems through interactions with key transcription factors. Here, we performed ChIP-Seq to identify target genes of DDX5 in cultured undifferentiated spermatogonia in order to elucidate its role in maintenance of the male germline.
Project description:DDX5, or PLZF co-immunoprecipitation in lysates from cultured undifferentiated spermatogonia followed by identification of eluted proteins using mass spectrometry. IgG control IPs included.
Project description:The maintenance of spermatogenesis in adult males is dependent on a population of mitotic germ cells with self-renewal potential known as undifferentiated spermatogonia. Regulation of undifferentiated spermatogonia function is dependent on transcriptional and post-transcriptional mechanisms. We have identified an essential role for the RNA helicase DDX5 in undifferentiated spermatogonia through generation of a UBC-CreERT2;Ddx5flox/flox mouse model that allows tamoxifen-dependent Ddx5 ablation. To identify genes regulated by DDX5, we generated lines of cultured undifferentiated spermatogonia from these mice and treated cells with tamoxifen (TMX) to induce Ddx5 knockout or vehicle (VEH) as a control, and performed RNA-sequencing analysis to compare these conditions.
Project description:Neural cell fate specification is a highly regulated process in which stem cells sequentially possess the transcriptional profile changed and give raise to particular cell lineages such as neuron and glial cells. The NTERA2 cell line is one of the well-characterized hEC cells that can be used as a model system for identifying the regulatory network during human pluripotent stem cells differentiated toward the neural lineage. It has been demonstrated that DDX5 and DDX17, as transcriptional co-regulator, can play a role in various cellular mechanisms depending on their interaction with the different types of transcription factors. Here, we performed ChIP-seq for DDX5 and DDX17 to identify their preferential binding targets during stage transition from pluripotent to neural cell lineage.