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Hybrid HIV/MSCV LTR enhances transgene expression of lentiviral vectors in human CD34(+) hematopoietic cells.
Stem Cells. 2001; 19(3):236-46.SC

Abstract

HIV-based lentiviral vectors can transduce nondividing cells, an important advantage over murine leukemia virus (MLV)-based vectors when transducing slowly dividing hematopoietic stem cells. However, we find that in human CD34(+) hematopoietic cells, the HIV-based vectors with an internal cytomegalovirus (CMV) promoter express transgenes 100- to 1,000-fold less than the MLV-based retroviral vector murine stem cell virus (MSCV). To increase the expression of the integrated lentivirus, we replaced CMV promoter with that of the Rous sarcoma virus or MSCV and obtained a modest augmentation in expression. A more dramatic effect was seen when the CMV enhancer/promoter was removed and the HIV long-terminal repeat (LTR) was replaced by a novel HIV/MSCV hybrid LTR. This vector retains the ability to transduce nondividing cells but now expresses its transgene (enhanced green fluorescent protein) 10- to 100-fold greater than the original HIV-based vector. When compared under identical conditions, the HIV vector with the hybrid LTR transduced a higher percentage of CD34(+) cells than the MSCV-based retroviral vector (19.4% versus 2.4%). The number of transduced cells and level of transgene expression remain constant over 5-8 weeks as determined by long-term culture-initiating cells, fluoresence-activated cell sorting, and nonobese diabetic/severe combined immunodeficiency repopulation assay.

Authors+Show Affiliations

Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia 19104, USA. jkchoi@mail.med.upenn.eduNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

11359949

Citation

Choi, J K., et al. "Hybrid HIV/MSCV LTR Enhances Transgene Expression of Lentiviral Vectors in Human CD34(+) Hematopoietic Cells." Stem Cells (Dayton, Ohio), vol. 19, no. 3, 2001, pp. 236-46.
Choi JK, Hoang N, Vilardi AM, et al. Hybrid HIV/MSCV LTR enhances transgene expression of lentiviral vectors in human CD34(+) hematopoietic cells. Stem Cells. 2001;19(3):236-46.
Choi, J. K., Hoang, N., Vilardi, A. M., Conrad, P., Emerson, S. G., & Gewirtz, A. M. (2001). Hybrid HIV/MSCV LTR enhances transgene expression of lentiviral vectors in human CD34(+) hematopoietic cells. Stem Cells (Dayton, Ohio), 19(3), 236-46.
Choi JK, et al. Hybrid HIV/MSCV LTR Enhances Transgene Expression of Lentiviral Vectors in Human CD34(+) Hematopoietic Cells. Stem Cells. 2001;19(3):236-46. PubMed PMID: 11359949.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Hybrid HIV/MSCV LTR enhances transgene expression of lentiviral vectors in human CD34(+) hematopoietic cells. AU - Choi,J K, AU - Hoang,N, AU - Vilardi,A M, AU - Conrad,P, AU - Emerson,S G, AU - Gewirtz,A M, PY - 2001/5/22/pubmed PY - 2001/7/28/medline PY - 2001/5/22/entrez SP - 236 EP - 46 JF - Stem cells (Dayton, Ohio) JO - Stem Cells VL - 19 IS - 3 N2 - HIV-based lentiviral vectors can transduce nondividing cells, an important advantage over murine leukemia virus (MLV)-based vectors when transducing slowly dividing hematopoietic stem cells. However, we find that in human CD34(+) hematopoietic cells, the HIV-based vectors with an internal cytomegalovirus (CMV) promoter express transgenes 100- to 1,000-fold less than the MLV-based retroviral vector murine stem cell virus (MSCV). To increase the expression of the integrated lentivirus, we replaced CMV promoter with that of the Rous sarcoma virus or MSCV and obtained a modest augmentation in expression. A more dramatic effect was seen when the CMV enhancer/promoter was removed and the HIV long-terminal repeat (LTR) was replaced by a novel HIV/MSCV hybrid LTR. This vector retains the ability to transduce nondividing cells but now expresses its transgene (enhanced green fluorescent protein) 10- to 100-fold greater than the original HIV-based vector. When compared under identical conditions, the HIV vector with the hybrid LTR transduced a higher percentage of CD34(+) cells than the MSCV-based retroviral vector (19.4% versus 2.4%). The number of transduced cells and level of transgene expression remain constant over 5-8 weeks as determined by long-term culture-initiating cells, fluoresence-activated cell sorting, and nonobese diabetic/severe combined immunodeficiency repopulation assay. SN - 1066-5099 UR - https://www.unboundmedicine.com/medline/citation/11359949/Hybrid_HIV/MSCV_LTR_enhances_transgene_expression_of_lentiviral_vectors_in_human_CD34_+__hematopoietic_cells_ L2 - https://doi.org/10.1634/stemcells.19-3-236 DB - PRIME DP - Unbound Medicine ER -