{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:19:41Z","timestamp":1788308381186,"version":"build-2803163510"},"reference-count":60,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2023,12,15]],"date-time":"2023-12-15T00:00:00Z","timestamp":1702598400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the European Regional Development Fund and the Republic of Cyprus through the Research and Innovation Foundation","award":["EXCELLENCE\/1216\/0092"],"award-info":[{"award-number":["EXCELLENCE\/1216\/0092"]}]},{"name":"the European Regional Development Fund and the Republic of Cyprus through the Research and Innovation Foundation","award":["EXCELLENCE\/0421\/0086"],"award-info":[{"award-number":["EXCELLENCE\/0421\/0086"]}]},{"name":"the Cyprus Institute of Neurology &amp; Genetics","award":["EXCELLENCE\/1216\/0092"],"award-info":[{"award-number":["EXCELLENCE\/1216\/0092"]}]},{"name":"the Cyprus Institute of Neurology &amp; Genetics","award":["EXCELLENCE\/0421\/0086"],"award-info":[{"award-number":["EXCELLENCE\/0421\/0086"]}]},{"name":"project \u201cNew Infrastructure for the Diagnosis and Therapy of Patients\u201d was funded by Norway Grants","award":["EXCELLENCE\/1216\/0092"],"award-info":[{"award-number":["EXCELLENCE\/1216\/0092"]}]},{"name":"project \u201cNew Infrastructure for the Diagnosis and Therapy of Patients\u201d was funded by Norway Grants","award":["EXCELLENCE\/0421\/0086"],"award-info":[{"award-number":["EXCELLENCE\/0421\/0086"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Therapy via the gene addition of the anti-sickling \u03b2AS3-globin transgene is potentially curative for all \u03b2-hemoglobinopathies and therefore of particular clinical and commercial interest. This study investigates GLOBE-based lentiviral vectors (LVs) for \u03b2AS3-globin addition and evaluates strategies for an increased \u03b2-like globin expression without vector dose escalation. First, we report the development of a GLOBE-derived LV, GLV2-\u03b2AS3, which, compared to its parental vector, adds anti-sickling action and a transcription-enhancing 848-bp transcription terminator element, retains high vector titers and allows for superior \u03b2-like globin expression in primary patient-derived hematopoietic stem and progenitor cells (HSPCs). Second, prompted by our previous correction of HBBIVSI\u2212110(G&gt;A) thalassemia based on RNApol(III)-driven shRNAs in mono- and combination therapy, we analyzed a series of novel LVs for the RNApol(II)-driven constitutive or late-erythroid expression of HBBIVSI\u2212110(G&gt;A)-specific miRNA30-embedded shRNAs (shRNAmiR). This included bifunctional LVs, allowing for concurrent \u03b2AS3-globin expression. LVs were initially compared for their ability to achieve high \u03b2-like globin expression in HBBIVSI\u2212110(G&gt;A)-transgenic cells, before the evaluation of shortlisted candidate LVs in HBBIVSI\u2212110(G&gt;A)-homozygous HSPCs. The latter revealed that \u03b2-globin promoter-driven designs for monotherapy with HBBIVSI\u2212110(G&gt;A)-specific shRNAmiRs only marginally increased \u03b2-globin levels compared to untransduced cells, whereas bifunctional LVs combining miR30-shRNA with \u03b2AS3-globin expression showed disease correction similar to that achieved by the parental GLV2-\u03b2AS3 vector. Our results establish the feasibility of high titers for LVs containing the full HBB transcription terminator, emphasize the importance of the HBB terminator for the high-level expression of HBB-like transgenes, qualify the therapeutic utility of late-erythroid HBBIVSI\u2212110(G&gt;A)-specific miR30-shRNA expression and highlight the exceptional potential of GLV2-\u03b2AS3 for the treatment of severe \u03b2-hemoglobinopathies.<\/jats:p>","DOI":"10.3390\/cells12242848","type":"journal-article","created":{"date-parts":[[2023,12,15]],"date-time":"2023-12-15T06:53:16Z","timestamp":1702623196000},"page":"2848","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Evaluation of Mono- and Bi-Functional GLOBE-Based Vectors for Therapy of \u03b2-Thalassemia by HBBAS3 Gene Addition and Mutation-Specific RNA Interference"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0438-8770","authenticated-orcid":false,"given":"Lola","family":"Koniali","sequence":"first","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina","family":"Flouri","sequence":"additional","affiliation":[{"name":"Gene Expression and Therapy Group, Department of Medical and Molecular Genetics, King\u2019s College London, Guy\u2019s Hospital, London SE1 9RT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markela I.","family":"Kostopoulou","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9456-5990","authenticated-orcid":false,"given":"Nikoletta Y.","family":"Papaioannou","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4410-3119","authenticated-orcid":false,"given":"Panayiota L.","family":"Papasavva","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Basma","family":"Naiisseh","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2546-6478","authenticated-orcid":false,"given":"Coralea","family":"Stephanou","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7808-0344","authenticated-orcid":false,"given":"Anthi","family":"Demetriadou","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Sitarou","sequence":"additional","affiliation":[{"name":"Thalassemia Clinic Larnaca, Larnaca General Hospital, 6301 Larnaca, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soteroula","family":"Christou","sequence":"additional","affiliation":[{"name":"Thalassemia Clinic Nicosia, Archbishop Makarios III Hospital, 1474 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3185-5314","authenticated-orcid":false,"given":"Michael N.","family":"Antoniou","sequence":"additional","affiliation":[{"name":"Gene Expression and Therapy Group, Department of Medical and Molecular Genetics, King\u2019s College London, Guy\u2019s Hospital, London SE1 9RT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0064-7034","authenticated-orcid":false,"given":"Marina","family":"Kleanthous","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8287-6724","authenticated-orcid":false,"given":"Petros","family":"Patsali","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3920-9584","authenticated-orcid":false,"given":"Carsten W.","family":"Lederer","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology & Genetics, 6 Iroon Avenue, 2371 Nicosia, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1111\/ijlh.13885","article-title":"The Hemoglobinopathies, Molecular Disease Mechanisms and Diagnostics","volume":"44","author":"Harteveld","year":"2022","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Halim-Fikri, B.H., Lederer, C.W., Baig, A.A., Mat-Ghani, S.N.A., Syed-Hassan, S.N.R.K., Yusof, W., Rashid, D.A., Azman, N.F., Fucharoen, S., and Panigoro, R. (2022). Global Globin Network Consensus Paper: Classification and Stratified Roadmaps for Improved Thalassaemia Care and Prevention in 32 Countries. J. Pers. Med., 12.","DOI":"10.3390\/jpm12040552"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1038\/nature09328","article-title":"Transfusion Independence and HMGA2 Activation after Gene Therapy of Human \u03b2-Thalassaemia","volume":"467","author":"Payen","year":"2010","journal-title":"Nature"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.1056\/NEJMoa1705342","article-title":"Gene Therapy in Patients with Transfusion-Dependent \u03b2-Thalassemia","volume":"378","author":"Thompson","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1038\/s41591-018-0301-6","article-title":"Intrabone Hematopoietic Stem Cell Gene Therapy for Adult and Pediatric Patients Affected by Transfusion-Dependent \u00df-Thalassemia","volume":"25","author":"Marktel","year":"2019","journal-title":"Nat. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/s41591-021-01650-w","article-title":"Long-Term Outcomes of Lentiviral Gene Therapy for the \u03b2-Hemoglobinopathies: The HGB-205 Trial","volume":"28","author":"Magrin","year":"2022","journal-title":"Nat. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1056\/NEJMoa2113206","article-title":"Betibeglogene Autotemcel Gene Therapy for Non\u2013\u03b20\/\u03b20 Genotype \u03b2-Thalassemia","volume":"386","author":"Locatelli","year":"2022","journal-title":"N. Engl. J. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4628","DOI":"10.1182\/blood-2019-125807","article-title":"Long-Term Clinical Outcomes of Lentiglobin Gene Therapy for Transfusion-Dependent \u03b2-Thalassemia in the Northstar (HGB-204) Study","volume":"134","author":"Kwiatkowski","year":"2019","journal-title":"Blood"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1182\/blood-2019-128482","article-title":"Northstar-3: Interim Results from a Phase 3 Study Evaluating Lentiglobin Gene Therapy in Patients with Transfusion-Dependent \u03b2-Thalassemia and Either a \u03b20 or IVS-I-110 Mutation at Both Alleles of the HBB Gene","volume":"134","author":"Lal","year":"2019","journal-title":"Blood"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3543","DOI":"10.1182\/blood-2019-126046","article-title":"Northstar-2: Updated Safety and Efficacy Analysis of Lentiglobin Gene Therapy in Patients with Transfusion-Dependent \u03b2-Thalassemia and Non-\u03b20\/\u03b20 Genotypes","volume":"134","author":"Thompson","year":"2019","journal-title":"Blood"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.stem.2021.01.001","article-title":"Hematopoietic-Stem-Cell-Targeted Gene-Addition and Gene-Editing Strategies for \u03b2-Hemoglobinopathies","volume":"28","author":"Drysdale","year":"2021","journal-title":"Cell Stem Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2455","DOI":"10.1073\/pnas.78.4.2455","article-title":"Base Substitution in an Intervening Sequence of a Beta+-Thalassemic Human Globin Gene","volume":"78","author":"Spritz","year":"1981","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2551","DOI":"10.1002\/j.1460-2075.1986.tb04534.x","article-title":"Beta Zero Thalassemia Caused by a Base Substitution That Creates an Alternative Splice Acceptor Site in an Intron","volume":"5","author":"Metherall","year":"1986","journal-title":"EMBO J."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Patsali, P., Papasavva, P., Christou, S., Sitarou, M., Antoniou, M.N., Lederer, C.W., and Kleanthous, M. (2020). Relative and Absolute Quantification of Aberrant and Normal Splice Variants in H\u0392BIVSI\u2212110(G>A) \u03b2-Thalassemia. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21186671"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e54","DOI":"10.1093\/nar\/gni052","article-title":"A GFP-Based Reporter System to Monitor Nonsense-Mediated mRNA Decay","volume":"33","author":"Paillusson","year":"2005","journal-title":"Nucleic Acids Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7399","DOI":"10.1074\/jbc.M512931200","article-title":"Humanized Beta-Thalassemia Mouse Model Containing the Common IVSI-110 Splicing Mutation","volume":"281","author":"Vadolas","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1182\/blood-2015-03-629618","article-title":"Plerixafor+G-CSF-Mobilized CD34+ Cells Represent an Optimal Graft Source for Thalassemia Gene Therapy","volume":"126","author":"Karponi","year":"2015","journal-title":"Blood"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1182\/blood.V114.22.978.978","article-title":"Following Beta-Globin Gene Transfer, the Production of Hemoglobin Depends Upon the Beta-Thalassemia Genotype","volume":"114","author":"Breda","year":"2009","journal-title":"Blood"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"8673","DOI":"10.1073\/pnas.90.18.8673","article-title":"Restoration of Correct Splicing in Thalassemic Pre-mRNA by Antisense Oligonucleotides","volume":"90","author":"Dominski","year":"1993","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","first-page":"700","article-title":"Restoration of Correct Splicing in IVSI-110 Mutation of \u03b2-Globin Gene with Antisense Oligonucleotides: Implications and Applications in Functional Assay Development","volume":"20","author":"Derakhshan","year":"2017","journal-title":"Iran. J. Basic Med. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1097\/MPH.0b013e3181639afe","article-title":"Correction of Aberrant Pre-mRNA Splicing by Antisense Oligonucleotides in Beta-Thalassemia Egyptian Patients with IVSI-110 Mutation","volume":"30","author":"Mostafa","year":"2008","journal-title":"J. Pediatr. Hematol. Oncol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e419","DOI":"10.3324\/haematol.2018.189357","article-title":"Short-Hairpin RNA against Aberrant HBBIVSI-110(G>A) mRNA Restores \u03b2-Globin Levels in a Novel Cell Model and Acts as Mono-and Combination Therapy for \u03b2-Thalassemia in Primary Hematopoietic Stem Cells","volume":"103","author":"Patsali","year":"2018","journal-title":"Haematologica"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.omtn.2023.03.012","article-title":"Novel Lentiviral Vectors for Gene Therapy of Sickle Cell Disease Combining Gene Addition and Gene Silencing Strategies","volume":"32","author":"Brusson","year":"2023","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1056\/NEJMoa2029392","article-title":"Post-Transcriptional Genetic Silencing of BCL11A to Treat Sickle Cell Disease","volume":"384","author":"Esrick","year":"2021","journal-title":"N. Engl. J. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1016\/j.hoc.2017.06.010","article-title":"Gene Therapy Approaches to Hemoglobinopathies","volume":"31","author":"Ferrari","year":"2017","journal-title":"Hematol. Oncol. Clin. N. Am."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/S0009-8981(01)00672-6","article-title":"Diagnosis of Hemoglobinopathies: Electrophoresis vs. HPLC","volume":"313","author":"Ou","year":"2001","journal-title":"Clin. Chim. Acta"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/j.molcel.2009.01.008","article-title":"Transcriptional Termination Enhances Protein Expression in Human Cells","volume":"33","author":"West","year":"2009","journal-title":"Mol. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/S1097-2765(00)80464-5","article-title":"Terminal Exon Definition Occurs Cotranscriptionally and Promotes Termination of RNA Polymerase II","volume":"3","author":"Dye","year":"1999","journal-title":"Mol. Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.omtm.2018.10.014","article-title":"Pre-Clinical Development of a Lentiviral Vector Expressing the Anti-Sickling \u03b2AS3 Globin for Gene Therapy for Sickle Cell Disease","volume":"11","author":"Poletti","year":"2018","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1089\/hum.2018.061","article-title":"Gene Therapy for Sickle Cell Disease: A Lentiviral Vector Comparison Study","volume":"29","author":"Urbinati","year":"2018","journal-title":"Hum. Gene Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.omtm.2018.07.012","article-title":"An Optimized Lentiviral Vector Efficiently Corrects the Human Sickle Cell Disease Phenotype","volume":"10","author":"Weber","year":"2018","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_32","unstructured":"(2023, November 29). Study Details|Stem Cell Gene Therapy for Sickle Cell Disease|ClinicalTrials.Gov, Available online: https:\/\/www.clinicaltrials.gov\/study\/NCT02247843."},{"key":"ref_33","unstructured":"(2023, November 29). Study Details|Safety and Efficacy of Gene Therapy of the Sickle Cell Disease by Transplantation of an Autologous CD34+ Enriched Cell Fraction That Contains CD34+ Cells Transduced Ex Vivo with the GLOBE1 Lentiviral Vector Expressing the \u03b2AS3 Globin Gene in Patients with Sickle Cell Disease (DREPAGLOBE)|ClinicalTrials.Gov, Available online: https:\/\/www.clinicaltrials.gov\/study\/NCT03964792."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"10547","DOI":"10.1073\/pnas.0711666105","article-title":"In Vivo Selection of Genetically Modified Erythroblastic Progenitors Leads to Long-Term Correction of \u03b2-Thalassemia","volume":"105","author":"Miccio","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"27518","DOI":"10.1074\/jbc.M402578200","article-title":"A Recombinant Human Hemoglobin with Anti-Sickling Properties Greater than Fetal Hemoglobin","volume":"279","author":"Levasseur","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"5421","DOI":"10.1111\/j.1742-4658.2006.05534.x","article-title":"Design of Expression Vectors for RNA Interference Based on miRNAs and RNA Splicing","volume":"273","author":"Du","year":"2006","journal-title":"FEBS J."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Antoniou, M. (1991). Induction of Erythroid-Specific Expression in Murine Erythroleukemia Cell Lines. J. Mol. Biol., 7.","DOI":"10.1385\/0-89603-178-0:421"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0092-8674(78)90082-X","article-title":"Human Alpha-Globin Gene Expression Following Chromosomal Dependent Gene Transfer into Mouse Erythroleukemia Cells","volume":"15","author":"Deisseroth","year":"1978","journal-title":"Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/j.jcyt.2016.11.007","article-title":"Suitability of Small Diagnostic Peripheral-Blood Samples for Cell-Therapy Studies","volume":"19","author":"Stephanou","year":"2017","journal-title":"Cytotherapy"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1038\/gt.2015.60","article-title":"Measurement of Lentiviral Vector Titre and Copy Number by Cross-Species Duplex Quantitative PCR","volume":"23","author":"Christodoulou","year":"2016","journal-title":"Gene. Ther."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1089\/hgtb.2017.190","article-title":"Rapid and Sensitive Assessment of Globin Chains for Gene and Cell Therapy of Hemoglobinopathies","volume":"29","author":"Loucari","year":"2018","journal-title":"Hum. Gene. Ther. Methods"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1834","DOI":"10.1261\/rna.1062908","article-title":"Minimizing Variables among Hairpin-Based RNAi Vectors Reveals the Potency of shRNAs","volume":"14","author":"Boudreau","year":"2008","journal-title":"RNA"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Lebbink, R.J., Lowe, M., Chan, T., Khine, H., Wang, X., and McManus, M.T. (2011). Polymerase II Promoter Strength Determines Efficacy of MicroRNA Adapted shRNAs. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0026213"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1465","DOI":"10.1038\/mt.2015.113","article-title":"miRNA-Embedded shRNAs for Lineage-Specific BCL11A Knockdown and Hemoglobin F Induction","volume":"23","author":"Guda","year":"2015","journal-title":"Mol. Ther."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3065","DOI":"10.1002\/j.1460-2075.1994.tb06605.x","article-title":"Mutagenesis of Retroviral Vectors Transducing Human Beta-Globin Gene and Beta-Globin Locus Control Region Derivatives Results in Stable Transmission of an Active Transcriptional Structure","volume":"13","author":"Leboulch","year":"1994","journal-title":"EMBO J."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Miccio, A., Poletti, V., Tiboni, F., Rossi, C., Antonelli, A., Mavilio, F., and Ferrari, G. (2011). The GATA1-HS2 Enhancer Allows Persistent and Position-Independent Expression of a \u03b2-Globin Transgene. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0027955"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1038\/nbt.3062","article-title":"Genomic Discovery of Potent Chromatin Insulators for Human Gene Therapy","volume":"33","author":"Liu","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3445","DOI":"10.1182\/blood-2004-04-1427","article-title":"Successful Correction of the Human Beta-Thalassemia Major Phenotype Using a Lentiviral Vector","volume":"104","author":"Puthenveetil","year":"2004","journal-title":"Blood"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1863","DOI":"10.1038\/sj.mt.6300259","article-title":"Improved Human Beta-Globin Expression from Self-Inactivating Lentiviral Vectors Carrying the Chicken Hypersensitive Site-4 (CHS4) Insulator Element","volume":"15","author":"Arumugam","year":"2007","journal-title":"Mol. Ther."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"15012","DOI":"10.1038\/mtm.2015.12","article-title":"The Human Ankyrin 1 Promoter Insulator Sustains Gene Expression in a \u03b2-Globin Lentiviral Vector in Hematopoietic Stem Cells","volume":"2","author":"Romero","year":"2015","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.1016\/j.ymthe.2020.12.036","article-title":"Lentiviral Vector ALS20 Yields High Hemoglobin Levels with Low Genomic Integrations for Treatment of Beta-Globinopathies","volume":"29","author":"Breda","year":"2021","journal-title":"Mol. Ther."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"4175","DOI":"10.1182\/blood-2007-08-108647","article-title":"Locus Control Region Elements HS1 and HS4 Enhance the Therapeutic Efficacy of Globin Gene Transfer in \u03b2-Thalassemic Mice","volume":"110","author":"Lisowski","year":"2007","journal-title":"Blood"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1038\/sj.mt.6300394","article-title":"Globin Lentiviral Vector Insertions Can Perturb the Expression of Endogenous Genes in Beta-Thalassemic Hematopoietic Cells","volume":"16","author":"Hargrove","year":"2008","journal-title":"Mol. Ther."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"2740","DOI":"10.3324\/haematol.2020.276634","article-title":"Inclusion of a Short Hairpin RNA Targeting BCL11A into a \u03b2-Globin Expressing Vector Allows Concurrent Synthesis of Curative Adult and Fetal Hemoglobin","volume":"106","author":"Lourenco","year":"2021","journal-title":"Haematologica"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2817","DOI":"10.1182\/blood-2010-08-300723","article-title":"Therapeutic Levels of Fetal Hemoglobin in Erythroid Progeny of \u03b2-Thalassemic CD34+ Cells after Lentiviral Vector-Mediated Gene Transfer","volume":"117","author":"Wilber","year":"2011","journal-title":"Blood"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"2841","DOI":"10.1016\/j.ymthe.2021.04.037","article-title":"Coordinated \u03b2-Globin Expression and \u03b12-Globin Reduction in a Multiplex Lentiviral Gene Therapy Vector for \u03b2-Thalassemia","volume":"29","author":"Nualkaew","year":"2021","journal-title":"Mol. Ther."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1038\/nbt1176","article-title":"A Genetic Strategy to Treat Sickle Cell Anemia by Coregulating Globin Transgene Expression and RNA Interference","volume":"24","author":"Samakoglu","year":"2006","journal-title":"Nat. Biotechnol."},{"key":"ref_58","first-page":"e59780","article-title":"Pooled CRISPR-Based Genetic Screens in Mammalian Cells","volume":"2019","author":"Chan","year":"2019","journal-title":"J. Vis. Exp."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1002\/emmm.201000083","article-title":"Correction of Beta-Thalassemia Major by Gene Transfer in Haematopoietic Progenitors of Pediatric Patients","volume":"2","author":"Roselli","year":"2010","journal-title":"EMBO Mol. Med."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Breda, L., Casu, C., Gardenghi, S., Bianchi, N., Cartegni, L., Narla, M., Yazdanbakhsh, K., Musso, M., Manwani, D., and Little, J. (2012). Therapeutic Hemoglobin Levels after Gene Transfer in \u03b2-Thalassemia Mice and in Hematopoietic Cells of \u03b2-Thalassemia and Sickle Cells Disease Patients. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0032345"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/24\/2848\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:39:28Z","timestamp":1760132368000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/24\/2848"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,15]]},"references-count":60,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["cells12242848"],"URL":"https:\/\/doi.org\/10.3390\/cells12242848","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,12,15]]}}}