{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T12:07:59Z","timestamp":1779365279193,"version":"3.53.0"},"posted":{"date-parts":[[2023,4,3]]},"group-title":"Neuroscience","reference-count":45,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,4,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>OBJECTIVE<\/jats:title>\n                  <jats:p>\n                    Spinocerebellar ataxia type 12 (SCA12) is a neurodegenerative disease caused by expansion of a CAG repeat in the\n                    <jats:italic>PPP2R2B gene<\/jats:italic>\n                    . Here we tested the hypothesis that the\n                    <jats:italic>PPP2R2B antisense<\/jats:italic>\n                    (\n                    <jats:italic>PPP2R2B-AS1<\/jats:italic>\n                    ) transcript containing a CUG repeat is expressed and contributes to SCA12 pathogenesis.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>METHODS<\/jats:title>\n                  <jats:p>\n                    Expression of\n                    <jats:italic>PPP2R2B-AS1<\/jats:italic>\n                    transcript was detected in SCA12 human induced pluripotent stem cells (iPSCs), iPSC-derived NGN2 neurons, and SCA12 knock-in mouse brains using strand-specific RT-PCR (SS-RT-PCR). The tendency of expanded\n                    <jats:italic>PPP2R2B-AS1<\/jats:italic>\n                    (\n                    <jats:italic>expPPP2R2B-AS1<\/jats:italic>\n                    ) RNA to form foci, a marker of toxic processes involving mutant RNAs, was examined in SCA12 cell models by fluorescence\n                    <jats:italic>in situ<\/jats:italic>\n                    hybridization. The toxic effect of\n                    <jats:italic>expPPP2R2B-AS1<\/jats:italic>\n                    transcripts on SK-N-MC neuroblastoma cells was evaluated by caspase 3\/7 activity. Western blot was used to examine the expression of repeat associated non-ATG-initiated (RAN) translation of\n                    <jats:italic>expPPP2R2B-AS1<\/jats:italic>\n                    transcript in SK-N-MC cells.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>RESULTS<\/jats:title>\n                  <jats:p>\n                    The repeat region in\n                    <jats:italic>PPP2R2B<\/jats:italic>\n                    gene locus is bidirectionally transcribed in SCA12 iPSCs, iPSC-derived NGN2 neurons, and SCA12 mouse brains. Transfected\n                    <jats:italic>expPPP2R2B-AS1<\/jats:italic>\n                    transcripts are toxic to SK-N-MC cells, and the toxicity may be mediated, at least in part, by the RNA secondary structure. The\n                    <jats:italic>expPPP2R2B-AS1<\/jats:italic>\n                    transcripts form CUG RNA foci in SK-N-MC cells.\n                    <jats:italic>expPPP2R2B-AS1<\/jats:italic>\n                    transcript is translated in the Alanine ORF via repeat-associated non-ATG (RAN) translation, which is diminished by single nucleotide interruptions within the CUG repeat, and MBNL1 overexpression.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>INTERPRETATION<\/jats:title>\n                  <jats:p>\n                    These findings suggest that\n                    <jats:italic>PPP2R2B-AS1<\/jats:italic>\n                    contributes to SCA12 pathogenesis, and may therefore provide a novel therapeutic target for the disease.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.04.02.535298","type":"posted-content","created":{"date-parts":[[2023,4,4]],"date-time":"2023-04-04T01:01:03Z","timestamp":1680570063000},"source":"Crossref","is-referenced-by-count":2,"title":["Bidirectional transcription at the\n                  <i>PPP2R2B<\/i>\n                  gene locus in spinocerebellar ataxia type 12"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0431-275X","authenticated-orcid":false,"given":"Chengqian","family":"Zhou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans B.","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fatemeh J.","family":"Bakhsh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingyao","family":"Ying","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Russell L.","family":"Margolis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pan P.","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2023040607451046000_2023.04.02.535298v1.1","first-page":"39","article-title":"Clinical Characterization of Genetically Diagnosed Cases of Spinocerebellar Ataxia Type 12 from India","volume":"5","year":"2017","journal-title":"Mov. 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