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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,8,12]]},"abstract":"<jats:p>\n                    Intronic GGGGCC hexanucleotide repeat expansions in\n                    <jats:italic toggle=\"yes\">C9orf72<\/jats:italic>\n                    are the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Despite its intronic location, this repeat avidly supports synthesis of pathogenic dipeptide repeat (DPR) proteins via repeat-associated non-AUG (RAN) translation. However, the template RNA species that undergoes RAN translation endogenously remains unclear. Using long-read based 5\u2032 RNA ligase-mediated rapid amplification of cDNA ends (5\u2032 Repeat-RLM-RACE), we identified\n                    <jats:italic toggle=\"yes\">C9orf72<\/jats:italic>\n                    transcripts initiating within intron 1 in a C9BAC mouse model, patient-derived iNeurons, and iNeuron-derived polysomes. These cryptic m\n                    <jats:sup>7<\/jats:sup>\n                    G-capped mRNAs are at least partially polyadenylated and are more abundant than transcripts derived from intron retention or circular intron lariats. In RAN translation reporter assays, intronic template transcripts\u2013even those with short (32 nucleotide) leaders\u2013exhibited robust expression compared to exon\u2013intron and repeat-containing lariat reporters. To assess endogenous repeat-containing lariat RNA contributions to RAN translation, we enhanced endogenous lariat stability by knocking down the lariat debranching enzyme Dbr1. However, this modulation did not impact DPR production in patient-derived iNeurons. These findings identify cryptic, linear, m\n                    <jats:sup>7<\/jats:sup>\n                    G-capped intron-initiating\n                    <jats:italic toggle=\"yes\">C9orf72<\/jats:italic>\n                    mRNAs as an endogenous template for RAN translation and DPR production, with implications for disease pathogenesis and therapeutic development.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2507334122","type":"journal-article","created":{"date-parts":[[2025,8,4]],"date-time":"2025-08-04T19:05:07Z","timestamp":1754334307000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":14,"title":["Cryptic intronic transcriptional initiation generates efficient endogenous mRNA templates for C9orf72-associated RAN translation"],"prefix":"10.1073","volume":"122","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3418-8067","authenticated-orcid":false,"given":"Shannon L.","family":"Miller","sequence":"first","affiliation":[{"name":"Department of Neurology, University of Michigan","place":["Ann Arbor"]},{"name":"Cellular and Molecular Biology Graduate Program, University of Michigan","place":["Ann Arbor"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5562-9920","authenticated-orcid":false,"given":"Katelyn M.","family":"Green","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of Michigan","place":["Ann Arbor"]},{"name":"Cellular and Molecular Biology Graduate Program, University of Michigan","place":["Ann Arbor"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4582-4137","authenticated-orcid":false,"given":"Bradley","family":"Crone","sequence":"additional","affiliation":[{"name":"Department of Computational Medicine and Bioinformatics, University of Michigan","place":["Ann Arbor"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica A.","family":"Switzenberg","sequence":"additional","affiliation":[{"name":"Department of Computational Medicine and Bioinformatics, University of Michigan","place":["Ann Arbor"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1965-2012","authenticated-orcid":false,"given":"Elizabeth M. 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