Long-term and efficient expression of human β-globin gene in a hematopoietic cell line using a new site-specific integrating non-viral system

Long-term and efficient expression of human β-globin gene in a hematopoietic cell line using a new site-specific integrating non-viral system

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ABSTRACT Targeted integration of a therapeutic gene at specific loci in safe genomic regions by a non-viral vector can restore the function of the damaged gene. This approach also minimizes


the potential genotoxic effects of transferred DNA. In this study, we have developed a non-viral vector that functions according to site-specific recombination (SSR). The vector contained a


bacterial backbone and puromycin resistance gene (_pur__r_), a β-globin expressing cassette and an _attB_ recombination site. We used phiC31 integrase to insert a copy of the vector into


specific genomic locations of a human hematopoietic cell line. Site-specific integration of the vector with one or two copies in the transcriptionally active regions of the genome was


confirmed. After genomic integration, we used Cre recombinase to remove the bacterial backbone and _pur__r_. This removal was verified by negative selection and genomic PCR screening.


Following deletion of these sequences, the stable β-chain expression was continued for several months in the absence of selective pressure. Consequently, this vector may potentially be a


powerful tool for _ex vivo_ correction of β-globinopathies such as β-thalassemia through successful genomic integration of a functional copy of the globin gene into the patient’s target


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support SIMILAR CONTENT BEING VIEWED BY OTHERS LENTIVIRAL GLOBIN GENE THERAPY WITH REDUCED-INTENSITY CONDITIONING IN ADULTS WITH Β-THALASSEMIA: A PHASE 1 TRIAL Article 03 January 2022 GENE


REPLACEMENT OF Α-GLOBIN WITH Β-GLOBIN RESTORES HEMOGLOBIN BALANCE IN Β-THALASSEMIA-DERIVED HEMATOPOIETIC STEM AND PROGENITOR CELLS Article 18 March 2021 THERAPEUTIC ADENINE BASE EDITING OF


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sincerest gratitude to Professor Michelle Calos for providing the pDB2 and pCMV/Int vectors. This study was funded by grants provided by Royan Institute. AUTHOR INFORMATION AUTHORS AND


AFFILIATIONS * Department of Pharmaceutical Biotechnology and Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical


Sciences, Isfahan, Iran K Dormiani, H Mir Mohammad Sadeghi & H Sadeghi-Aliabadi * Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for


Biotechnology, ACECR, Isfahan, Iran K Ghaedi, M Forouzanfar & M H Nasr-Esfahani * Biology Department, School of Sciences, University of Isfahan, Isfahan, Iran K Ghaedi * Department of


Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran H Baharvand * Department of Developmental


Biology, University of Science and Culture, ACECR, Tehran, Iran H Baharvand Authors * K Dormiani View author publications You can also search for this author inPubMed Google Scholar * H Mir


Mohammad Sadeghi View author publications You can also search for this author inPubMed Google Scholar * H Sadeghi-Aliabadi View author publications You can also search for this author


inPubMed Google Scholar * K Ghaedi View author publications You can also search for this author inPubMed Google Scholar * M Forouzanfar View author publications You can also search for this


author inPubMed Google Scholar * H Baharvand View author publications You can also search for this author inPubMed Google Scholar * M H Nasr-Esfahani View author publications You can also


search for this author inPubMed Google Scholar CORRESPONDING AUTHORS Correspondence to H Baharvand or M H Nasr-Esfahani. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no


conflict of interest. ADDITIONAL INFORMATION Supplementary Information accompanies this paper on Gene Therapy website SUPPLEMENTARY INFORMATION SUPPLEMENTARY FIGURE LEGENDS (DOC 26 KB)


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Sadeghi, H., Sadeghi-Aliabadi, H. _et al._ Long-term and efficient expression of human β-globin gene in a hematopoietic cell line using a new site-specific integrating non-viral system.


_Gene Ther_ 22, 663–674 (2015). https://doi.org/10.1038/gt.2015.30 Download citation * Received: 24 September 2014 * Revised: 07 March 2015 * Accepted: 16 March 2015 * Published: 01 April


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