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Performance- and safety-enhanced lentiviral vectors containing the human interferon-beta scaffold attachment region and the chicken beta-globin insulator.
Blood. 2003 Jun 15; 101(12):4717-24.Blood

Abstract

Retroviral vectors are the most efficient means of stable gene delivery to hematopoietic stem cells (HSCs). However, transgene expression from retroviral vectors is frequently subject to the negative influence of chromosomal sequences flanking the site of integration. Toward the development of autonomous transgene expression cassettes, we inserted the human interferon-beta scaffold attachment region (IFN-SAR) and the chicken beta-globin 5' DNase I hypersensitive site 4 (5'HS4) insulator both separately and together into a series of self-inactivating (SIN) lentiviral vector backbones. Transduced cells of the human CD34+ hematopoietic progenitor line KG1a-pooled populations as well as individual clones harboring single integrants--were analyzed for reporter expression during culture periods of up to 4 months. Vectors carrying both the 5'HS4 insulator and the IFN-SAR consistently outperformed control vectors without inserts as well as vectors carrying either element alone. The performance of a set of vectors containing the murine stem cell virus long terminal repeat as an internal promoter was subsequently assessed during in vitro monocytic differentiation of transduced primary human CD34+ cord blood cells. Similar to what was observed in the KG1a hematopoietic progenitor cell model, optimal reporter expression in primary monocytes was obtained with the vector bearing both regulatory elements. These findings indicate that the 5'HS4/IFN-SAR combination is particularly effective at maintaining open chromatin domains permissive for high-level transgene expression at early and late stages of hematopoietic development, and thus could be of utility in HSC-directed retroviral vector-mediated gene transfer applications.

Authors+Show Affiliations

Hematopoiesis Department, Flow Cytometry Facility, American Red Cross, Rockville, MD 20855, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

12586614

Citation

Ramezani, Ali, et al. "Performance- and Safety-enhanced Lentiviral Vectors Containing the Human Interferon-beta Scaffold Attachment Region and the Chicken Beta-globin Insulator." Blood, vol. 101, no. 12, 2003, pp. 4717-24.
Ramezani A, Hawley TS, Hawley RG. Performance- and safety-enhanced lentiviral vectors containing the human interferon-beta scaffold attachment region and the chicken beta-globin insulator. Blood. 2003;101(12):4717-24.
Ramezani, A., Hawley, T. S., & Hawley, R. G. (2003). Performance- and safety-enhanced lentiviral vectors containing the human interferon-beta scaffold attachment region and the chicken beta-globin insulator. Blood, 101(12), 4717-24.
Ramezani A, Hawley TS, Hawley RG. Performance- and Safety-enhanced Lentiviral Vectors Containing the Human Interferon-beta Scaffold Attachment Region and the Chicken Beta-globin Insulator. Blood. 2003 Jun 15;101(12):4717-24. PubMed PMID: 12586614.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Performance- and safety-enhanced lentiviral vectors containing the human interferon-beta scaffold attachment region and the chicken beta-globin insulator. AU - Ramezani,Ali, AU - Hawley,Teresa S, AU - Hawley,Robert G, Y1 - 2003/02/13/ PY - 2003/2/15/pubmed PY - 2004/2/5/medline PY - 2003/2/15/entrez SP - 4717 EP - 24 JF - Blood JO - Blood VL - 101 IS - 12 N2 - Retroviral vectors are the most efficient means of stable gene delivery to hematopoietic stem cells (HSCs). However, transgene expression from retroviral vectors is frequently subject to the negative influence of chromosomal sequences flanking the site of integration. Toward the development of autonomous transgene expression cassettes, we inserted the human interferon-beta scaffold attachment region (IFN-SAR) and the chicken beta-globin 5' DNase I hypersensitive site 4 (5'HS4) insulator both separately and together into a series of self-inactivating (SIN) lentiviral vector backbones. Transduced cells of the human CD34+ hematopoietic progenitor line KG1a-pooled populations as well as individual clones harboring single integrants--were analyzed for reporter expression during culture periods of up to 4 months. Vectors carrying both the 5'HS4 insulator and the IFN-SAR consistently outperformed control vectors without inserts as well as vectors carrying either element alone. The performance of a set of vectors containing the murine stem cell virus long terminal repeat as an internal promoter was subsequently assessed during in vitro monocytic differentiation of transduced primary human CD34+ cord blood cells. Similar to what was observed in the KG1a hematopoietic progenitor cell model, optimal reporter expression in primary monocytes was obtained with the vector bearing both regulatory elements. These findings indicate that the 5'HS4/IFN-SAR combination is particularly effective at maintaining open chromatin domains permissive for high-level transgene expression at early and late stages of hematopoietic development, and thus could be of utility in HSC-directed retroviral vector-mediated gene transfer applications. SN - 0006-4971 UR - https://www.unboundmedicine.com/medline/citation/12586614/Performance__and_safety_enhanced_lentiviral_vectors_containing_the_human_interferon_beta_scaffold_attachment_region_and_the_chicken_beta_globin_insulator_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-4971(20)50631-9 DB - PRIME DP - Unbound Medicine ER -