Project description:The aim of the experiment is to reveal splicing isoforms induced by transient expression of Fritillaria borealis U2AF in HEK293T cells. Cells were transfected with different combinations of U2AF and U2AF2 subunits paralogues. A cycloheximide treatment was applied to inhibit nonsense-mediated decay and to stabilize transcripts produced by aberrant splicing. After cDNA conversion, target isoforms were amplified and sequenced.
Project description:The goal of the experiment is to reveal RNA binding preferences of Fritillaria borealis U2AF2 paralogues, using CLIP-seq. Coding sequences of U2AF2alpha, U2AF2beta and Env negative control were fused to a C-terminal V5 tag. Protein expression constructs were transfected in mammalian cell culture. After cross-linking, RNA-protein complexes were recovered with anti-V5 magnetic beads.
