{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T13:31:35Z","timestamp":1791293495314,"version":"4.1.0"},"reference-count":47,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2016,10,30]],"date-time":"2016-10-30T00:00:00Z","timestamp":1477785600000},"content-version":"vor","delay-in-days":29,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004887","name":"Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences","doi-asserted-by":"publisher","award":["Grant\/Award Number: 92\u201003\u201087\u201021886"],"award-info":[{"award-number":["Grant\/Award Number: 92\u201003\u201087\u201021886"]}],"id":[{"id":"10.13039\/501100004887","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Journal of Gene Medicine"],"published-print":{"date-parts":[[2016,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>\u03b2\u2010thalassemia comprises a major group of human genetic disorders involving a decrease in or an end to the normal synthesis of the \u03b2\u2010globin chains of hemoglobin. KLF1 is a key regulatory molecule involved in the \u03b3\u2010 to \u03b2\u2010globin gene switching process directly inducing the expression of the \u03b2\u2010globin gene and indirectly repressing \u03b3\u2010globin. The present study aimed to investigate the ability of an engineered CRISPR\/<jats:italic>Cas9<\/jats:italic> system with respect to disrupting the <jats:italic>KLF1<\/jats:italic> gene to inhibit the \u03b3\u2010 to \u03b2\u2010hemoglobin switching process in K562 cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We targeted three sites on the <jats:italic>KLF1<\/jats:italic> gene, two of which are upstream of codon 288 in exon 2 and the other site being in exon 3.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The average indel percentage in the cells transfected with CRISPR a, b and c was approximately 24%. Relative quantification was performed for the assessment of \u03b3\u2010globin expression. The levels of \u03b3\u2010globin mRNA on day 5 of differentiation were 8.1\u2010, 7.7\u2010 and 1.8\u2010fold in the cells treated with CRISPR\/<jats:italic>Cas9<\/jats:italic> a, b and c, respectively,compared to untreated cells. The measurement of HbF expression levels confirmed the same results.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The findings obtained in the present study support the induction of an indel mutation in the <jats:italic>KLF1<\/jats:italic> gene leading to a null allele. As a result, the effect of <jats:italic>KLF1<\/jats:italic> on the expression of <jats:italic>BCL11A<\/jats:italic> is decreased and its inhibitory effect on \u03b3\u2010globin gene expression is removed. Application of CRISPR technology to induce an indel in the <jats:italic>KLF1<\/jats:italic> gene in adult erythroid progenitors may provide a method for activating fetal hemoglobin expression in individuals with \u03b2\u2010thalassemia or sickle cell disease.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jgm.2928","type":"journal-article","created":{"date-parts":[[2016,9,26]],"date-time":"2016-09-26T17:27:02Z","timestamp":1474910822000},"page":"294-301","source":"Crossref","is-referenced-by-count":48,"title":["Genetic disruption of the <i>KLF1<\/i> gene to overexpress the \u03b3\u2010globin gene using the CRISPR\/<i>Cas9<\/i> system"],"prefix":"10.1002","volume":"18","author":[{"given":"Laleh","family":"Shariati","sequence":"first","affiliation":[{"name":"Department of Genetics and Molecular Biology, School of Medicine Isfahan University of Medical Sciences  Isfahan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hossein","family":"Khanahmad","sequence":"additional","affiliation":[{"name":"Department of Genetics and Molecular Biology, School of Medicine Isfahan University of Medical Sciences  Isfahan Iran"},{"name":"Pediatric Inherited Diseases Research Center, Research Institute for Primordial Prevention of Non\u2010communicable Disease Isfahan University of Medical Sciences  Isfahan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mansoor","family":"Salehi","sequence":"additional","affiliation":[{"name":"Department of Genetics and Molecular Biology, School of Medicine Isfahan University of Medical Sciences  Isfahan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zahra","family":"Hejazi","sequence":"additional","affiliation":[{"name":"Department of Genetics and Molecular Biology, School of Medicine Isfahan University of Medical Sciences  Isfahan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ilnaz","family":"Rahimmanesh","sequence":"additional","affiliation":[{"name":"Department of Genetics and Molecular Biology, School of Medicine Isfahan University of Medical Sciences  Isfahan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad Amin","family":"Tabatabaiefar","sequence":"additional","affiliation":[{"name":"Department of Genetics and Molecular Biology, School of Medicine Isfahan University of Medical Sciences  Isfahan Iran"},{"name":"Pediatric Inherited Diseases Research Center, Research Institute for Primordial Prevention of Non\u2010communicable Disease Isfahan University of Medical Sciences  Isfahan Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad Hossein","family":"Modarressi","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, School of Medicine Tehran University of Medical Sciences  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2016,10,30]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.4103\/0976-9668.136245"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.2174\/156652408786241393"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.hoc.2010.08.009"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1537-2995.2012.03837.x"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMcibr1010767"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2010-11-316893"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(86)92205-1"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(72)92592-5"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1097\/00000441-194804000-00008"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00439-009-0770-2"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/35066048"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199406093302303"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2009-04-217901"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.79.14.4428"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI111464"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199505183322001"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199506153322404"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2003-02-0478"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0022838"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.5483\/BMBRep.2009.42.8.493"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature08243"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1165409"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2006-02-004184"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng.637"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng.630"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2006-01-008912"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1018384108"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.exphem.2012.04.007"},{"key":"e_1_2_6_30_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-03-331371"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1097\/MOH.0b013e32835f59ba"},{"key":"e_1_2_6_32_1","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.110.120717"},{"key":"e_1_2_6_33_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.3.1156"},{"key":"e_1_2_6_34_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1258096"},{"key":"e_1_2_6_35_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1225829"},{"key":"e_1_2_6_36_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.24733"},{"key":"e_1_2_6_37_1","doi-asserted-by":"publisher","DOI":"10.1101\/gr.145441.112"},{"key":"e_1_2_6_38_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10529-016-2101-8"},{"key":"e_1_2_6_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2012.23"},{"key":"e_1_2_6_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1521-6926(04)00068-4"},{"key":"e_1_2_6_41_1","doi-asserted-by":"publisher","DOI":"10.1182\/asheducation-2011.1.459"},{"key":"e_1_2_6_42_1","first-page":"000","article-title":"Gamma reactivation using the spongy effect of KLF1\u2010binding site sequence: an approach in gene therapy for beta thalassemia","volume":"19","author":"Heydari N","year":"2016","journal-title":"J Basic Med Sci"},{"key":"e_1_2_6_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms8085"},{"key":"e_1_2_6_44_1","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1897310"},{"key":"e_1_2_6_45_1","first-page":"131","article-title":"Analyzing inhibition of BCL11A gene expression in K562 cells by RNAi","volume":"2","author":"Vikas U","year":"2013","journal-title":"J BioSci Biotechnol"},{"key":"e_1_2_6_46_1","unstructured":"KL McCance SE Huether 2015"},{"key":"e_1_2_6_47_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1231143"},{"key":"e_1_2_6_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2623"}],"container-title":["The Journal of Gene Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjgm.2928","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jgm.2928","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,26]],"date-time":"2023-09-26T12:00:22Z","timestamp":1695729622000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jgm.2928"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,10]]},"references-count":47,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2016,10]]}},"alternative-id":["10.1002\/jgm.2928"],"URL":"https:\/\/doi.org\/10.1002\/jgm.2928","archive":["Portico"],"relation":{},"ISSN":["1099-498X","1521-2254"],"issn-type":[{"value":"1099-498X","type":"print"},{"value":"1521-2254","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,10]]}}}