Alt-RPL36 downregulates the PI3K-AKT-mTOR signaling pathway by interacting with TMEM24

We characterize non-annotated protein alt-RPL36 which is encoded by RPL36 transcript isoform 2 and overlaps with the RPL36 open reading frame. Its function is examined by specific CRISPR deletion of the GTG start codon located 5' to the RPL36 ATG initiation site. Overall changes in transcript expression are evaluated by comparison with wild type HEK293T cells. RPL36 protein expression is not affected by the altRPL36 start codon deletion. Overall design: Gene expression changes between two biological replicates of control (HEK293T) cells and HEK293T cells with alt-RPL36 knockout are quantified. The 200 kb DNA region around CRISPR/Cas9 target sites in our alt-RPL36 knockout and knock-in HEK293T cell lines was enriched using Samplix Xdrop Technology and sequenced via long-read Oxford Nanopore Technology.