This study aims to modify and apply STARR-seq (self-transcribing active regulatory region sequencing) to quantitatively map whole-genome enhancer activities in the present or absent of Zelda. With integrative analysis and meta-analysis of pre-established related datasets, we will be able to discover the bona fide signatures of ZGA enhancer networks, and reconstruct the regulome of ZGA. I have successfully performed NGS-based functional genomic approaches combined with big data analysis (including RNA-seq, transcription factor ChIP-seq, pol II ChIP-seq, MNase-seq), and have published papers in journals with high impact factors. In addition, I have the supports from and communications with the expertise on the approaches used in my aims, including Alexander Stark at IMP, Vienna. I have also finished the most onerous and rate-limiting step which is to generate genomic library for STARR-seq. There, I would expect to accomplish this work. Aim 1. To delineate the chromatin accessibility landscape of Zelda expressing S2 cells. Aim 2. To map quantitative genome-wide Zelda-dependent enhancers activity in S2 cells by STARR-seq. Aim3. To reconstruct the regulome in S2 cells reprogramed by Zelda.
