{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T04:08:22Z","timestamp":1790482102574,"version":"4.1.0"},"reference-count":24,"publisher":"National Academy of Sciences","issue":"15","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2006,4,11]]},"abstract":"\n A single nucleotide polymorphism (SNP) in the sickle \u03b2-globin gene (\u03b2\n S<\/jats:sup>\n ) leads to sickle cell anemia. Sickling increases sharply with deoxy sickle Hb concentration and decreases with increasing fetal \u03b3-globin concentration. Measures that decrease sickle Hb concentration should have an antisickling effect. RNA interference (RNAi) uses small interfering (si)RNAs for sequence-specific gene silencing. A \u03b2\n S<\/jats:sup>\n siRNA with position 10 of the guide strand designed to align with the targeted \u03b2\n \n S<\/jats:italic>\n <\/jats:sup>\n SNP specifically silences \u03b2\n \n S<\/jats:italic>\n <\/jats:sup>\n gene expression without affecting the expression of the \u03b3-globin or normal \u03b2-globin (\u03b2\n \n A<\/jats:italic>\n <\/jats:sup>\n ) genes. Silencing is increased by altering the 5\u2032 end of the siRNA antisense (guide) strand to enhance its binding to the RNA-induced silencing complex (RISC). Specific \u03b2\n S<\/jats:sup>\n silencing was demonstrated by using a luciferase reporter and full-length \u03b2\n S<\/jats:sup>\n cDNA transfected into HeLa cells and mouse erythroleukemia cells, where it was expressed in the context of the endogenous \u03b2-globin gene promoter and the locus control region enhancers. When this strategy was used to target \u03b2\n \n E<\/jats:italic>\n <\/jats:sup>\n , silencing was not limited to the mutant gene but also targeted the normal \u03b2\n \n A<\/jats:italic>\n <\/jats:sup>\n gene. siRNAs, mismatched with their target at position 10, guided mRNA cleavage in all cases except when two bulky purines were aligned. The specific silencing of the \u03b2\n \n S<\/jats:italic>\n <\/jats:sup>\n -globin gene, as compared with \u03b2\n \n E<\/jats:italic>\n <\/jats:sup>\n , as well as studies of silencing SNP mutants in other diseases, indicates that siRNAs developed to target a disease-causing SNP will be specific if the mutant residue is a pyrimidine and the normal residue is a purine.\n <\/jats:p>","DOI":"10.1073\/pnas.0601309103","type":"journal-article","created":{"date-parts":[[2006,4,3]],"date-time":"2006-04-03T20:35:00Z","timestamp":1144096500000},"page":"5953-5958","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":45,"title":["Determinants of specific RNA interference-mediated silencing of human \u03b2-globin alleles differing by a single nucleotide polymorphism"],"prefix":"10.1073","volume":"103","author":[{"given":"Derek M.","family":"Dykxhoorn","sequence":"first","affiliation":[{"name":"*CBR Institute for Biomedical Research and Department of Pediatrics, Harvard Medical School, Boston, MA 02115;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa D.","family":"Schlehuber","sequence":"additional","affiliation":[{"name":"*CBR Institute for Biomedical Research and Department of Pediatrics, Harvard Medical School, Boston, MA 02115;"},{"name":"Harvard\u2013Massachusetts Institute of Technology Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irving M.","family":"London","sequence":"additional","affiliation":[{"name":"Department of Biology and"},{"name":"Harvard\u2013Massachusetts Institute of Technology Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judy","family":"Lieberman","sequence":"additional","affiliation":[{"name":"*CBR Institute for Biomedical Research and Department of Pediatrics, Harvard Medical School, Boston, MA 02115;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2006,4,11]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(04)17192-4"},{"key":"e_1_3_3_2_2","first-page":"577","volume-title":"Disorders of Hemoglobin: Genetics, Pathophysiology, and Clinical Management","author":"Ferrone F.","year":"2001","unstructured":"F. 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