In 2016, Stefanie Gerstberger completed her doctoral thesis at The Rockefeller University titled "A Census of Human RNA-Binding Proteins and Characterization of the DEDDH RNA Exonuclease NEF-SP in the 3' End Maturation of 28S Ribosomal RNA." The thesis focused on two main areas: the comprehensive cataloging of human RNA-binding proteins (RBPs) and the characterization of the RNA exonuclease NEF-SP in the processing of 28S ribosomal RNA. The first part of the thesis involved creating a detailed census of 1,542 manually curated RBPs. This census included categorizing RBPs based on their interactions with different classes of RNA, analyzing their evolutionary patterns, abundance, and tissue-specific expression. The study also explored co-regulated gene expression during developmental processes to gain insights into regulatory pathways. The findings suggested that classifying RBPs into their main regulatory RNA pathways could help understand the disease phenotypes of proteins involved in the same RNA metabolic pathways. The thesis also identified a novel RBP, FAM98A, which is rich in RG/GG motifs and binds to G-rich regions in mRNA targets. The study demonstrated that FAM98A does not affect target mRNA stability when knocked down, indicating its potential role in RNA metabolism. In the second part of the thesis, Gerstberger focused on the uncharacterized, highly tissue-specific RNA exonuclease NEF-sp. She characterized its function in the processing of pre-28S ribosomal RNA. The study revealed that NEF-sp, a DEDDh RNA exonuclease, is involved in the removal of the 3' external transcribed spacer (3'ETS) of the large 47S rRNA precursor in Drosophila melanogaster. The findings showed that NEF-sp is highly expressed in gonads and that its absence leads to gonad development arrest. The study demonstrated that exonucleolytic trimming is essential for 28S rRNA maturation in higher eukaryotes and that the expression of a factor involved in a core RNA metabolic process can be highly regulated. The research suggested an additional level of posttranscriptional gene regulation in the maturation of 28S rRNA, mediated by the regulated expression of RNA exonucleases. The thesis also included acknowledgments to the mentors, colleagues, and institutions that supported the research. The work was conducted under the mentorship of Thomas Tuschl and involved collaboration with various labs and facilities. The study utilized a combination of computational methods, experimental approaches, and advanced sequencing technologies to analyze RNA-binding proteins and their roles in RNA metabolism. The findings contribute to a better understanding of posttranscriptional gene regulation and the role of RBPs in human diseases.In 2016, Stefanie Gerstberger completed her doctoral thesis at The Rockefeller University titled "A Census of Human RNA-Binding Proteins and Characterization of the DEDDH RNA Exonuclease NEF-SP in the 3' End Maturation of 28S Ribosomal RNA." The thesis focused on two main areas: the comprehensive cataloging of human RNA-binding proteins (RBPs) and the characterization of the RNA exonuclease NEF-SP in the processing of 28S ribosomal RNA. The first part of the thesis involved creating a detailed census of 1,542 manually curated RBPs. This census included categorizing RBPs based on their interactions with different classes of RNA, analyzing their evolutionary patterns, abundance, and tissue-specific expression. The study also explored co-regulated gene expression during developmental processes to gain insights into regulatory pathways. The findings suggested that classifying RBPs into their main regulatory RNA pathways could help understand the disease phenotypes of proteins involved in the same RNA metabolic pathways. The thesis also identified a novel RBP, FAM98A, which is rich in RG/GG motifs and binds to G-rich regions in mRNA targets. The study demonstrated that FAM98A does not affect target mRNA stability when knocked down, indicating its potential role in RNA metabolism. In the second part of the thesis, Gerstberger focused on the uncharacterized, highly tissue-specific RNA exonuclease NEF-sp. She characterized its function in the processing of pre-28S ribosomal RNA. The study revealed that NEF-sp, a DEDDh RNA exonuclease, is involved in the removal of the 3' external transcribed spacer (3'ETS) of the large 47S rRNA precursor in Drosophila melanogaster. The findings showed that NEF-sp is highly expressed in gonads and that its absence leads to gonad development arrest. The study demonstrated that exonucleolytic trimming is essential for 28S rRNA maturation in higher eukaryotes and that the expression of a factor involved in a core RNA metabolic process can be highly regulated. The research suggested an additional level of posttranscriptional gene regulation in the maturation of 28S rRNA, mediated by the regulated expression of RNA exonucleases. The thesis also included acknowledgments to the mentors, colleagues, and institutions that supported the research. The work was conducted under the mentorship of Thomas Tuschl and involved collaboration with various labs and facilities. The study utilized a combination of computational methods, experimental approaches, and advanced sequencing technologies to analyze RNA-binding proteins and their roles in RNA metabolism. The findings contribute to a better understanding of posttranscriptional gene regulation and the role of RBPs in human diseases.