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Efficiency of different fragment lengths of the ubiquitous chromatin opening element HNRPA2B1-CBX3 in driving human CD18 gene expression within self-inactivating lentiviral vectors for gene therapy applications.

Abstract

Patients with leukocyte adhesion deficiency type 1 (LAD1) suffer from life-threatening bacterial infections due to mutations in the common β2 integrin subunit (CD18/ITGB2 gene). We tested different fragments of the ubiquitous chromatin opening element (UCOE) from the human HNRPA2B1-CBX3 locus for their efficiency in driving the human CD18 gene expression and compared it with that of an elongation factor 1 alpha promoter (EF1αL, 1169 bp; EF1αS 248 bp) and a murine stem cell virus (MSCV) promoter within the context of the same lentiviral vector backbone. These vectors were tested in vitro for the human CD18 gene expression on the surface of CD34+ hematopoietic stem cells (HSCs) isolated from both moderate and severe LAD1 patients. Among the promoters tested in the patients' CD34+ HSCs, only U631 bp, U652 bp, U1262 bp, 5' 2.2 kb A2UCOE and EF1αS resulted in higher percentage of CD18+CD34+ cells comparable to that of the MSCV promoter. The U655 bp, U723 bp, U1296 bp, U2598 bp and EF1αL promoters resulted in comparatively lower numbers of CD18+CD34+ cells. This study would be useful in investigating the human CD18 gene expression in an ex vivo experiment to demonstrate the phenotypic correction of LAD1 in a pre-clinical model.

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  • Authors+Show Affiliations

    ,

    Gene Therapy Laboratory, Department of Integrative Biology, School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore 632 014, Tamil Nadu, India.

    ,

    Gene Therapy Laboratory, Department of Integrative Biology, School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore 632 014, Tamil Nadu, India.

    ,

    Grow Research Laboratory, Narayana Nethralaya Foundation, Bangalore 560 099, Karnataka, India.

    Gene Therapy Laboratory, Department of Integrative Biology, School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore 632 014, Tamil Nadu, India. Electronic address: everette.nelson@vit.ac.in.

    Source

    Gene 710: 2019 Aug 20 pg 265-272

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    31200085

    Citation

    Gopinath, Chitra, et al. "Efficiency of Different Fragment Lengths of the Ubiquitous Chromatin Opening Element HNRPA2B1-CBX3 in Driving Human CD18 Gene Expression Within Self-inactivating Lentiviral Vectors for Gene Therapy Applications." Gene, vol. 710, 2019, pp. 265-272.
    Gopinath C, Chodisetty S, Ghosh A, et al. Efficiency of different fragment lengths of the ubiquitous chromatin opening element HNRPA2B1-CBX3 in driving human CD18 gene expression within self-inactivating lentiviral vectors for gene therapy applications. Gene. 2019;710:265-272.
    Gopinath, C., Chodisetty, S., Ghosh, A., & Nelson, E. J. R. (2019). Efficiency of different fragment lengths of the ubiquitous chromatin opening element HNRPA2B1-CBX3 in driving human CD18 gene expression within self-inactivating lentiviral vectors for gene therapy applications. Gene, 710, pp. 265-272. doi:10.1016/j.gene.2019.06.016.
    Gopinath C, et al. Efficiency of Different Fragment Lengths of the Ubiquitous Chromatin Opening Element HNRPA2B1-CBX3 in Driving Human CD18 Gene Expression Within Self-inactivating Lentiviral Vectors for Gene Therapy Applications. Gene. 2019 Aug 20;710:265-272. PubMed PMID: 31200085.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Efficiency of different fragment lengths of the ubiquitous chromatin opening element HNRPA2B1-CBX3 in driving human CD18 gene expression within self-inactivating lentiviral vectors for gene therapy applications. AU - Gopinath,Chitra, AU - Chodisetty,Sarvani, AU - Ghosh,Arkasubhra, AU - Nelson,Everette Jacob Remington, Y1 - 2019/06/11/ PY - 2019/02/16/received PY - 2019/06/06/revised PY - 2019/06/10/accepted PY - 2019/6/15/pubmed PY - 2019/6/15/medline PY - 2019/6/15/entrez KW - CD18 KW - Gene therapy KW - Hematopoietic stem cells (HSCs) KW - Lentiviral vectors KW - Leukocyte adhesion deficiency type 1 (LAD1) KW - Ubiquitous chromatin opening element (UCOE) SP - 265 EP - 272 JF - Gene JO - Gene VL - 710 N2 - Patients with leukocyte adhesion deficiency type 1 (LAD1) suffer from life-threatening bacterial infections due to mutations in the common β2 integrin subunit (CD18/ITGB2 gene). We tested different fragments of the ubiquitous chromatin opening element (UCOE) from the human HNRPA2B1-CBX3 locus for their efficiency in driving the human CD18 gene expression and compared it with that of an elongation factor 1 alpha promoter (EF1αL, 1169 bp; EF1αS 248 bp) and a murine stem cell virus (MSCV) promoter within the context of the same lentiviral vector backbone. These vectors were tested in vitro for the human CD18 gene expression on the surface of CD34+ hematopoietic stem cells (HSCs) isolated from both moderate and severe LAD1 patients. Among the promoters tested in the patients' CD34+ HSCs, only U631 bp, U652 bp, U1262 bp, 5' 2.2 kb A2UCOE and EF1αS resulted in higher percentage of CD18+CD34+ cells comparable to that of the MSCV promoter. The U655 bp, U723 bp, U1296 bp, U2598 bp and EF1αL promoters resulted in comparatively lower numbers of CD18+CD34+ cells. This study would be useful in investigating the human CD18 gene expression in an ex vivo experiment to demonstrate the phenotypic correction of LAD1 in a pre-clinical model. SN - 1879-0038 UR - https://www.unboundmedicine.com/medline/citation/31200085/Efficiency_of_different_fragment_lengths_of_the_ubiquitous_chromatin_opening_element_HNRPA2B1-CBX3_in_driving_human_CD18_gene_expression_within_self-inactivating_lentiviral_vectors_for_gene_therapy_applications L2 - https://linkinghub.elsevier.com/retrieve/pii/S0378-1119(19)30576-1 DB - PRIME DP - Unbound Medicine ER -