RT Journal Article SR Electronic T1 The RNA-binding profile of the splicing factor SRSF6 in immortalized human pancreatic β-cells JF Life Science Alliance JO Life Sci. Alliance FD Life Science Alliance LLC SP e202000825 DO 10.26508/lsa.202000825 VO 4 IS 3 A1 Maria Inês Alvelos A1 Mirko Brüggemann A1 FX Reymond Sutandy A1 Jonàs Juan-Mateu A1 Maikel Luis Colli A1 Anke Busch A1 Miguel Lopes A1 Ângela Castela A1 Annemieke Aartsma-Rus A1 Julian König A1 Kathi Zarnack A1 Décio L Eizirik YR 2021 UL https://www.life-science-alliance.org/content/4/3/e202000825.abstract AB In pancreatic β-cells, the expression of the splicing factor SRSF6 is regulated by GLIS3, a transcription factor encoded by a diabetes susceptibility gene. SRSF6 down-regulation promotes β-cell demise through splicing dysregulation of central genes for β-cells function and survival, but how RNAs are targeted by SRSF6 remains poorly understood. Here, we define the SRSF6 binding landscape in the human pancreatic β-cell line EndoC-βH1 by integrating individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP) under basal conditions with RNA sequencing after SRSF6 knockdown. We detect thousands of SRSF6 bindings sites in coding sequences. Motif analyses suggest that SRSF6 specifically recognizes a purine-rich consensus motif consisting of GAA triplets and that the number of contiguous GAA triplets correlates with increasing binding site strength. The SRSF6 positioning determines the splicing fate. In line with its role in β-cell function, we identify SRSF6 binding sites on regulated exons in several diabetes susceptibility genes. In a proof-of-principle, the splicing of the susceptibility gene LMO7 is modulated by antisense oligonucleotides. Our present study unveils the splicing regulatory landscape of SRSF6 in immortalized human pancreatic β-cells.