{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T01:31:04Z","timestamp":1781659864528,"version":"3.54.5"},"reference-count":28,"publisher":"Springer Science and Business Media LLC","issue":"12","license":[{"start":{"date-parts":[[2019,9,9]],"date-time":"2019-09-09T00:00:00Z","timestamp":1567987200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2019,9,9]],"date-time":"2019-09-09T00:00:00Z","timestamp":1567987200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["81571448"],"award-info":[{"award-number":["81571448"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31872800"],"award-info":[{"award-number":["31872800"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Foundation of Guangzhou Science and Technology Commission","award":["201803040009"],"award-info":[{"award-number":["201803040009"]}]},{"DOI":"10.13039\/501100003453","name":"Natural Science Foundation of Guangdong Province","doi-asserted-by":"crossref","award":["2014A030312012"],"award-info":[{"award-number":["2014A030312012"]}],"id":[{"id":"10.13039\/501100003453","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Guangdong Province Science and Technology Project","award":["2016B030229008"],"award-info":[{"award-number":["2016B030229008"]}]},{"name":"Youth Project by Education of Guangdong Province","award":["2015KQNCX132\/B"],"award-info":[{"award-number":["2015KQNCX132\/B"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Ann Hematol"],"published-print":{"date-parts":[[2019,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:p>Haemoglobin (Hb) H-constant spring (CS) alpha thalassaemia (- -\/-\u03b1<jats:sup>CS<\/jats:sup>) is the most common type of nondeletional Hb H disease in southern China. The CRISPR\/Cas9-based gene correction of patient-specific induced pluripotent stem cells (iPSCs) and cell transplantation now represent a therapeutic solution for this genetic disease. We designed primers for the target sites using CRISPR\/Cas9 to specifically edit the HBA2 gene with an Hb-CS mutation. After applying a correction-specific PCR assay to purify the corrected clones followed by sequencing to confirm the mutation correction, we verified that the purified clones retained full pluripotency and exhibited a normal karyotype. This strategy may be promising in the future, although it is far from representing a solution for the treatment of HbH-CS thalassemia now.<\/jats:p>","DOI":"10.1007\/s00277-019-03763-2","type":"journal-article","created":{"date-parts":[[2019,9,9]],"date-time":"2019-09-09T03:30:21Z","timestamp":1567999821000},"page":"2661-2671","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["CRISPR\/Cas9 gene correction of HbH-CS thalassemia-induced pluripotent stem cells"],"prefix":"10.1007","volume":"98","author":[{"given":"Xie","family":"Yingjun","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xie","family":"Yuhuan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chen","family":"Yuchang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Dongzhi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wang","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Song","family":"Bing","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Yi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu","family":"Dian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue","family":"Yanting","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiong","family":"Zeyu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liu","family":"Nengqing","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chen","family":"Diyu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4960-2112","authenticated-orcid":false,"given":"Sun","family":"Xiaofang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,9,9]]},"reference":[{"key":"3763_CR1","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1186\/1750-1172-5-13","volume":"5","author":"CL Harteveld","year":"2010","unstructured":"Harteveld CL, Higgs DR (2010) Alpha-thalassaemia. Orphanet J Rare Dis 5:13","journal-title":"Orphanet J Rare Dis"},{"issue":"5","key":"3763_CR2","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1097\/MPH.0000000000001160","volume":"40","author":"P Komvilaisak","year":"2018","unstructured":"Komvilaisak P, Jetsrisuparb A, Fucharoen G, Komwilaisak R, Jirapradittha J, Kiatchoosakun P (2018) Clinical course of homozygous hemoglobin constant spring in pediatric patients. J Pediatr Hematol Oncol 40(5):409\u2013412","journal-title":"J Pediatr Hematol Oncol"},{"issue":"8","key":"3763_CR3","doi-asserted-by":"publisher","first-page":"544","DOI":"10.1056\/NEJM200008243430804","volume":"343","author":"FE Chen","year":"2000","unstructured":"Chen FE, Ooi C, Ha SY, Cheung BM, Todd D, Liang R et al (2000) Genetic and clinical features of hemoglobin H disease in Chinese patients. N Engl J Med 343(8):544\u2013550","journal-title":"N Engl J Med"},{"issue":"16","key":"3763_CR4","doi-asserted-by":"publisher","first-page":"1479","DOI":"10.1056\/NEJMoa1705342","volume":"378","author":"AA Thompson","year":"2018","unstructured":"Thompson AA, Walters MC, Kwiatkowski J, Rasko JEJ, Ribeil JA, Hongeng S, Magrin E, Schiller GJ, Payen E, Semeraro M, Moshous D, Lefrere F, Puy H, Bourget P, Magnani A, Caccavelli L, Diana JS, Suarez F, Monpoux F, Brousse V, Poirot C, Brouzes C, Meritet JF, Pondarr\u00e9 C, Beuzard Y, Chr\u00e9tien S, Lefebvre T, Teachey DT, Anurathapan U, Ho PJ, von Kalle C, Kletzel M, Vichinsky E, Soni S, Veres G, Negre O, Ross RW, Davidson D, Petrusich A, Sandler L, Asmal M, Hermine O, de Montalembert M, Hacein-Bey-Abina S, Blanche S, Leboulch P, Cavazzana M (2018) Gene therapy in patients with transfusion-dependent beta-thalassemia. N Engl J Med 378(16):1479\u20131493","journal-title":"N Engl J Med"},{"issue":"1","key":"3763_CR5","doi-asserted-by":"publisher","first-page":"230","DOI":"10.1038\/mt.2011.195","volume":"20","author":"E Yannaki","year":"2012","unstructured":"Yannaki E, Papayannopoulou T, Jonlin E, Zervou F, Karponi G, Xagorari A, Becker P, Psatha N, Batsis I, Kaloyannidis P, Tahynopoulou V, Constantinou V, Bouinta A, Kotta K, Athanassiadou A, Anagnostopoulos A, Fassas A, Stamatoyannopoulos G (2012) Hematopoietic stem cell mobilization for gene therapy of adult patients with severe beta-thalassemia: results of clinical trials using G-CSF or plerixafor in splenectomized and nonsplenectomized subjects. Mol Ther 20(1):230\u2013238","journal-title":"Mol Ther"},{"issue":"1","key":"3763_CR6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1089\/hum.2006.151","volume":"18","author":"M Sadelain","year":"2007","unstructured":"Sadelain M, Boulad F, Galanello R, Giardina P, Locatelli F, Maggio A, Rivella S, Riviere I, Tisdale J (2007) Therapeutic options for patients with severe beta-thalassemia: the need for globin gene therapy. Hum Gene Ther 18(1):1\u20139","journal-title":"Hum Gene Ther"},{"key":"3763_CR7","first-page":"8631432","volume":"2018","author":"J Liu","year":"2018","unstructured":"Liu J, Higgins CA, Whitehouse JC, Harris SJ, Crawford H, Christiano AM et al (2018) Hair follicle dermal cells support expansion of murine and human embryonic and induced pluripotent stem cells and promote haematopoiesis in mouse cultures. Stem Cells Int 2018:8631432","journal-title":"Stem Cells Int"},{"issue":"6","key":"3763_CR8","doi-asserted-by":"publisher","first-page":"971","DOI":"10.1111\/bjh.14491","volume":"176","author":"MA Razaq","year":"2017","unstructured":"Razaq MA, Taylor S, Roberts DJ, Carpenter L (2017) A molecular roadmap of definitive erythropoiesis from human induced pluripotent stem cells. Br J Haematol 176(6):971\u2013983","journal-title":"Br J Haematol"},{"issue":"1575","key":"3763_CR9","doi-asserted-by":"publisher","first-page":"2274","DOI":"10.1098\/rstb.2011.0017","volume":"366","author":"JJ Unternaehrer","year":"2011","unstructured":"Unternaehrer JJ, Daley GQ (2011) Induced pluripotent stem cells for modelling human diseases. Philos Trans R Soc Lond Ser B Biol Sci 366(1575):2274\u20132285","journal-title":"Philos Trans R Soc Lond Ser B Biol Sci"},{"key":"3763_CR10","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1016\/j.omtn.2016.11.010","volume":"6","author":"YL Liu","year":"2017","unstructured":"Liu YL, Yang Y, Kang XJ, Lin B, Yu Q, Song B, Gao G, Chen Y, Sun X, Li X, Bu L, Fan Y (2017) One-step biallelic and scarless correction of a beta-thalassemia mutation in patient-specific iPSCs without drug selection. Mol Ther-Nucl Acids 6:57\u201367","journal-title":"Mol Ther-Nucl Acids"},{"issue":"4","key":"3763_CR11","doi-asserted-by":"publisher","first-page":"1164","DOI":"10.1182\/blood.V93.4.1164","volume":"93","author":"G Lucarelli","year":"1999","unstructured":"Lucarelli G, Clift RA, Galimberti M, Angelucci E, Giardini C, Baronciani D, Polchi P, Andreani M, Gaziev D, Erer B, Ciaroni A, D'Adamo F, Albertini F, Muretto P (1999) Bone marrow transplantation in adult thalassemic patients. Blood. 93(4):1164\u20131167","journal-title":"Blood."},{"issue":"2","key":"3763_CR12","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1093\/bib\/bbs017","volume":"14","author":"H Thorvaldsdottir","year":"2013","unstructured":"Thorvaldsdottir H, Robinson JT, Mesirov JP (2013) Integrative genomics viewer (IGV): high-performance genomics data visualization and exploration. Brief Bioinform 14(2):178\u2013192","journal-title":"Brief Bioinform"},{"issue":"1","key":"3763_CR13","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1038\/nbt.1754","volume":"29","author":"JT Robinson","year":"2011","unstructured":"Robinson JT, Thorvaldsdottir H, Winckler W, Guttman M, Lander ES, Getz G et al (2011) Integrative genomics viewer. Nat Biotechnol 29(1):24\u201326","journal-title":"Nat Biotechnol"},{"issue":"32","key":"3763_CR14","doi-asserted-by":"publisher","first-page":"16576","DOI":"10.1074\/jbc.M116.719237","volume":"291","author":"XH Niu","year":"2016","unstructured":"Niu XH, He WY, Song B, Ou ZH, Fan D, Chen YC, Fan Y, Sun X (2016) Combining single strand oligodeoxynucleotides and CRISPR\/Cas9 to correct gene mutations in beta-thalassemia-induced pluripotent stem cells. J Biol Chem 291(32):16576\u201316585","journal-title":"J Biol Chem"},{"issue":"2","key":"3763_CR15","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1002\/(SICI)1096-8652(199610)53:2<81::AID-AJH5>3.0.CO;2-#","volume":"53","author":"AC Kattamis","year":"1996","unstructured":"Kattamis AC, Camaschella C, Sivera P, Surrey S, Fortina P (1996) Human alpha-thalassemia syndromes: detection of molecular defects. Am J Hematol 53(2):81\u201391","journal-title":"Am J Hematol"},{"issue":"9","key":"3763_CR16","doi-asserted-by":"publisher","first-page":"1041","DOI":"10.1007\/s00439-016-1691-5","volume":"135","author":"YY Jang","year":"2016","unstructured":"Jang YY, Ye ZH (2016) Gene correction in patient-specific iPSCs for therapy development and disease modeling. Hum Genet 135(9):1041\u20131058","journal-title":"Hum Genet"},{"issue":"3","key":"3763_CR17","doi-asserted-by":"publisher","first-page":"570","DOI":"10.1038\/mt.2014.226","volume":"23","author":"C Smith","year":"2015","unstructured":"Smith C, Abalde-Atristain L, He C, Brodsky BR, Braunstein EM, Chaudhari P, Jang YY, Cheng L, Ye Z (2015) Efficient and allele-specific genome editing of disease loci in human iPSCs. Mol Ther 23(3):570\u2013577","journal-title":"Mol Ther"},{"issue":"3","key":"3763_CR18","doi-asserted-by":"publisher","first-page":"194","DOI":"10.4103\/0975-7406.99020","volume":"4","author":"JV Raja","year":"2012","unstructured":"Raja JV, Rachchh MA, Gokani RH (2012) Recent advances in gene therapy for thalassemia. J Pharm Bioallied Sci 4(3):194\u2013201","journal-title":"J Pharm Bioallied Sci"},{"issue":"7629","key":"3763_CR19","doi-asserted-by":"publisher","first-page":"384","DOI":"10.1038\/nature20134","volume":"539","author":"DP Dever","year":"2016","unstructured":"Dever DP, Bak RO, Reinisch A, Camarena J, Washington G, Nicolas CE, Pavel-Dinu M, Saxena N, Wilkens AB, Mantri S, Uchida N, Hendel A, Narla A, Majeti R, Weinberg KI, Porteus MH (2016) CRISPR\/Cas9 beta-globin gene targeting in human haematopoietic stem cells. Nature. 539(7629):384\u2013389","journal-title":"Nature."},{"issue":"6121","key":"3763_CR20","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1126\/science.1231143","volume":"339","author":"L Cong","year":"2013","unstructured":"Cong L, Ran FA, Cox D, Lin SL, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F (2013) Multiplex genome engineering using CRISPR\/Cas systems. Science. 339(6121):819\u2013823","journal-title":"Science."},{"issue":"8","key":"3763_CR21","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1038\/nbt.1927","volume":"29","author":"D Hockemeyer","year":"2011","unstructured":"Hockemeyer D, Wang HY, Kiani S, Lai CS, Gao Q, Cassady JP, Cost GJ, Zhang L, Santiago Y, Miller JC, Zeitler B, Cherone JM, Meng X, Hinkley SJ, Rebar EJ, Gregory PD, Urnov FD, Jaenisch R (2011) Genetic engineering of human pluripotent cells using TALE nucleases. Nat Biotechnol 29(8):731\u2013734","journal-title":"Nat Biotechnol"},{"issue":"9","key":"3763_CR22","doi-asserted-by":"publisher","first-page":"851","DOI":"10.1038\/nbt.1562","volume":"27","author":"D Hockemeyer","year":"2009","unstructured":"Hockemeyer D, Soldner F, Beard C, Gao Q, Mitalipova M, DeKelver RC et al (2009) Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27(9):851\u2013U110","journal-title":"Nat Biotechnol"},{"issue":"19","key":"3763_CR23","doi-asserted-by":"publisher","first-page":"3906","DOI":"10.1182\/blood-2012-03-420703","volume":"120","author":"CJ Chang","year":"2012","unstructured":"Chang CJ, Bouhassira EE (2012) Zinc-finger nuclease-mediated correction of alpha-thalassemia in iPS cells. Blood. 120(19):3906\u20133914","journal-title":"Blood."},{"issue":"48","key":"3763_CR24","doi-asserted-by":"publisher","first-page":"34671","DOI":"10.1074\/jbc.M113.496174","volume":"288","author":"N Ma","year":"2013","unstructured":"Ma N, Liao BJ, Zhang H, Wang LL, Shan YL, Xue YT, Huang K, Chen S, Zhou X, Chen Y, Pei D, Pan G (2013) Transcription activator-like effector nuclease (TALEN)-mediated gene correction in integration-free beta-thalassemia induced pluripotent stem cells. J Biol Chem 288(48):34671\u201334679","journal-title":"J Biol Chem"},{"issue":"19","key":"3763_CR25","doi-asserted-by":"publisher","first-page":"12079","DOI":"10.1074\/jbc.M114.624999","volume":"290","author":"N Ma","year":"2015","unstructured":"Ma N, Shan Y, Liao B, Kong G, Wang C, Huang K, Zhang H, Cai X, Chen S, Pei D, Chen N, Pan G (2015) Factor-induced reprogramming and zinc finger nuclease-aided gene targeting cause different genome instability in beta-thalassemia induced pluripotent stem cells (iPSCs). J Biol Chem 290(19):12079\u201312089","journal-title":"J Biol Chem"},{"issue":"9","key":"3763_CR26","doi-asserted-by":"publisher","first-page":"1526","DOI":"10.1101\/gr.173427.114","volume":"24","author":"F Xie","year":"2014","unstructured":"Xie F, Ye L, Chang JC, Beyer AI, Wang JM, Muench MO, Kan YW (2014) Seamless gene correction of beta-thalassemia mutations in patient-specific iPSCs using CRISPR\/Cas9 and piggyBac. Genome Res 24(9):1526\u20131533","journal-title":"Genome Res"},{"issue":"7660","key":"3763_CR27","doi-asserted-by":"publisher","first-page":"686","DOI":"10.1038\/nature23012","volume":"546","author":"H Kilpinen","year":"2017","unstructured":"Kilpinen H, Goncalves A, Leha A, Afzal V, Alasoo K, Ashford S, Bala S, Bensaddek D, Casale FP, Culley OJ, Danecek P, Faulconbridge A, Harrison PW, Kathuria A, McCarthy D, McCarthy SA, Meleckyte R, Memari Y, Moens N, Soares F, Mann A, Streeter I, Agu CA, Alderton A, Nelson R, Harper S, Patel M, White A, Patel SR, Clarke L, Halai R, Kirton CM, Kolb-Kokocinski A, Beales P, Birney E, Danovi D, Lamond AI, Ouwehand WH, Vallier L, Watt FM, Durbin R, Stegle O, Gaffney DJ (2017) Common genetic variation drives molecular heterogeneity in human iPSCs (vol 546, pg 370, 2017). Nature. 546(7660):686","journal-title":"Nature."},{"issue":"11","key":"3763_CR28","doi-asserted-by":"publisher","first-page":"1182","DOI":"10.1038\/nbt.3387","volume":"33","author":"AM Tsankov","year":"2015","unstructured":"Tsankov AM, Akopian V, Pop R, Chetty S, Gifford CA, Daheron L, Tsankova NM, Meissner A (2015) A qPCR ScoreCard quantifies the differentiation potential of human pluripotent stem cells. Nat Biotechnol 33(11):1182\u20131192","journal-title":"Nat Biotechnol"}],"container-title":["Annals of Hematology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00277-019-03763-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s00277-019-03763-2\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00277-019-03763-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,3]],"date-time":"2021-09-03T18:13:59Z","timestamp":1630692839000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s00277-019-03763-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,9,9]]},"references-count":28,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2019,12]]}},"alternative-id":["3763"],"URL":"https:\/\/doi.org\/10.1007\/s00277-019-03763-2","relation":{},"ISSN":["0939-5555","1432-0584"],"issn-type":[{"value":"0939-5555","type":"print"},{"value":"1432-0584","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,9,9]]},"assertion":[{"value":"20 January 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 July 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 September 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Compliance with ethical standards"}},{"value":"The authors declare that they have no conflict of interest.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"The experimental procedures and animal conditions were approved by the Animal Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}}]}}