{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T02:18:32Z","timestamp":1773368312197,"version":"3.50.1"},"reference-count":19,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T00:00:00Z","timestamp":1666224000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Ministry of Science and Higher Education of the Russian Federation","award":["075-15-2021-1058"],"award-info":[{"award-number":["075-15-2021-1058"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>The elevation of SMN transcript and protein level remains the principal aim of SMA therapy. Still, there is no standard molecular biomarker for the assessment of its efficacy. In the current study, we tested three methods of SMN transcript level measurement using real-time RT-PCR, quantitative fluorescent RT-PCR, and a semiquantitative RT-PCR gel densitometric assay. We examined several potential mRNA-based biomarkers and examined their sensitivity and reliability by comparing the obtained values in peripheral blood mononuclear cells of SMA patients, SMA carriers, and healthy individuals. We found that the mean percentage of full-length (FL-SMN) transcripts relative to the total sum of FL-SMN and exon 7-deleted (\u03947 SMN) transcripts detected by semiquantitative and quantitative fluorescence RT-PCR differed significantly between the three analyzed groups. The relevance of this biomarker was proven in an SMN2-targeting therapeutic experiment. We showed that the values of the biomarker changed significantly in SMA fibroblast cell cultures after treatment with therapeutic antisense oligonucleotides targeting the ISS-N1 site in intron 7 of the SMN2 gene. The obtained results indicate the convenience of using the mean percentage of FL-SMN transcripts determined by semiquantitative and quantitative fluorescence RT-PCR as a putative biomarker for the assessment of SMA therapy efficacy in vitro.<\/jats:p>","DOI":"10.3390\/genes13101911","type":"journal-article","created":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T20:35:55Z","timestamp":1666298155000},"page":"1911","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Evaluation of Mean Percentage of Full-Length SMN Transcripts as a Molecular Biomarker of Spinal Muscular Atrophy"],"prefix":"10.3390","volume":"13","author":[{"given":"Marianna","family":"Maretina","sequence":"first","affiliation":[{"name":"Department of Genomic Medicine, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint-Petersburg, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna","family":"Egorova","sequence":"additional","affiliation":[{"name":"Department of Genomic Medicine, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint-Petersburg, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0749-0079","authenticated-orcid":false,"given":"Kristina","family":"Lanko","sequence":"additional","affiliation":[{"name":"Department of Clinical Genetics, Erasmus MC University Medical Center, Dr. Molewaterplein Street 40, 3015 GD Rotterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vladislav","family":"Baranov","sequence":"additional","affiliation":[{"name":"Department of Genomic Medicine, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint-Petersburg, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2487-2423","authenticated-orcid":false,"given":"Anton","family":"Kiselev","sequence":"additional","affiliation":[{"name":"Department of Genomic Medicine, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, Mendeleevskaya Line 3, 199034 Saint-Petersburg, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/0092-8674(95)90460-3","article-title":"Identification and characterization of a spinal muscular atrophy-determining gene","volume":"80","author":"Lefebvre","year":"1995","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1093\/hmg\/8.7.1177","article-title":"A single nucleotide difference that alters splicing patterns distinguishes the SMA gene SMN1 from the copy gene SMN","volume":"8","author":"Monani","year":"1999","journal-title":"Hum. Mol. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Zheleznyakova, G.Y., Kiselev, A.V., Vakharlovsky, V.G., Rask-Andersen, M., Chavan, R., Egorova, A.A., Schi\u00f6th, H.B., and Baranov, V.S. (2011). Genetic and expression studies of SMN2 gene in Russian patients with spinal muscular atrophy type II and III. BMC Med. Genet., 12.","DOI":"10.1186\/1471-2350-12-96"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"339","DOI":"10.2174\/1389202919666180101154916","article-title":"Molecular Factors Involved in Spinal Muscular Atrophy Pathways as Possible Disease-modifying Candidates","volume":"19","author":"Maretina","year":"2018","journal-title":"Curr. Genomics"},{"key":"ref_5","first-page":"87","article-title":"Current Trends in the Diagnosis, Screening and Treatment of Spinal Muscular Atrophy","volume":"77","author":"Maretina","year":"2022","journal-title":"Ann. Russ. Acad. Med. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1723","DOI":"10.1056\/NEJMoa1702752","article-title":"Nusinersen versus Sham Control in Infantile-Onset Spinal Muscular Atrophy","volume":"377","author":"Finkel","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"842","DOI":"10.1016\/j.nmd.2019.09.007","article-title":"Nusinersen initiated in infants during the presymptomatic stage of spinal muscular atrophy: Interim efficacy and safety results from the Phase 2 NURTURE study","volume":"29","author":"Bertini","year":"2019","journal-title":"Neuromuscul. Disord."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1002\/mus.27363","article-title":"Expert recommendations and clinical considerations in the use of onasemnogene abeparvovec gene therapy for spinal muscular atrophy","volume":"64","author":"Kichula","year":"2021","journal-title":"Muscle Nerve"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"341","DOI":"10.4161\/rna.6.3.8723","article-title":"A short antisense oligonucleotide masking a unique intronic motif prevents skipping of a critical exon in spinal muscular atrophy","volume":"6","author":"Singh","year":"2009","journal-title":"RNA Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1634","DOI":"10.1101\/gad.1941310","article-title":"Antisense correction of SMN2 splicing in the CNS rescues necrosis in a type III SMA mouse model","volume":"24","author":"Hua","year":"2010","journal-title":"Genes Dev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"970","DOI":"10.1002\/ana.20836","article-title":"In vivo activation of SMN in spinal muscular atrophy carriers and patients treated with valproate","volume":"59","author":"Brichta","year":"2006","journal-title":"Ann. Neurol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1038\/ejhg.2009.116","article-title":"SMN transcript levels in leukocytes of SMA patients determined by absolute real-time PCR","volume":"18","author":"Tiziano","year":"2010","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Crawford, T.O., Paushkin, S.V., Kobayashi, D.T., Forrest, S.J., Joyce, C.L., Finkel, R.S., Kaufmann, P., Swoboda, K.J., Tiziano, D., and Lomastro, R. (2012). Evaluation of SMN protein, transcript, and copy number in the biomarkers for spinal muscular atrophy (BforSMA) clinical study. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0033572"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1038\/sj.ejhg.5201885","article-title":"A sensitive assay for measuring SMN mRNA levels in peripheral blood and in muscle samples of patients affected with spinal muscular atrophy","volume":"15","author":"Vezain","year":"2007","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1007\/s00439-006-0245-7","article-title":"Novel aminoglycosides increase SMN levels in spinal muscular atrophy fibroblasts","volume":"120","author":"Mattis","year":"2006","journal-title":"Hum. Genet."},{"key":"ref_16","first-page":"70","article-title":"Neural differentiation of patient-specific induced pluripotent stem cells from patients with a hereditary form of spinal muscular atrophy","volume":"11","author":"Valetdinova","year":"2016","journal-title":"Genes Cells"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1677\/jme.0.0250169","article-title":"Absolute quantification of mrna using real-time reverse transcription polymerase chain reaction assays","volume":"25","author":"Bustin","year":"2000","journal-title":"J. Mol. Endocrinol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1701","DOI":"10.1038\/s41591-021-01483-7","article-title":"Biodistribution of onasemnogene abeparvovec DNA, mRNA and SMN protein in human tissue","volume":"27","author":"Thomsen","year":"2021","journal-title":"Nat. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1056\/NEJMoa2009965","article-title":"Risdiplam in Type 1 Spinal Muscular Atrophy","volume":"384","author":"Baranello","year":"2021","journal-title":"N. Engl. J. Med."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/13\/10\/1911\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:58:31Z","timestamp":1760144311000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/13\/10\/1911"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,20]]},"references-count":19,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["genes13101911"],"URL":"https:\/\/doi.org\/10.3390\/genes13101911","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,20]]}}}