Design of Ad5F35 vectors for coordinated dual gene expression in candidate human hematopoietic stem cells.
(2010) In Experimental Hematology Apr 8. p.446-452- Abstract
- OBJECTIVE: Adenoviral vector mediated gene expression is an attractive approach to manipulate or report gene expression in human hematopoietic stem cells (HSC), when transient gene expression is preferred. Previous studies have demonstrated that fiber retargeted Ad5F35 vectors can mediate efficient gene transfer into human HSCs. In this study, we have investigated the potential of bi-directional promoter controlled Ad5F35 vector for coordinated dual gene expression in candidate HSCs. MATERIAL AND METHODS: We have engineered Ad5F35-DeltaLNGFR-BiDp encoding kinase domain deleted low-affinity NGF receptor (DeltaLNGFR) and green fluorescent protein (GFP) expression cassette controlled by a synthetic bi-directional promoter, which is composed... (More)
- OBJECTIVE: Adenoviral vector mediated gene expression is an attractive approach to manipulate or report gene expression in human hematopoietic stem cells (HSC), when transient gene expression is preferred. Previous studies have demonstrated that fiber retargeted Ad5F35 vectors can mediate efficient gene transfer into human HSCs. In this study, we have investigated the potential of bi-directional promoter controlled Ad5F35 vector for coordinated dual gene expression in candidate HSCs. MATERIAL AND METHODS: We have engineered Ad5F35-DeltaLNGFR-BiDp encoding kinase domain deleted low-affinity NGF receptor (DeltaLNGFR) and green fluorescent protein (GFP) expression cassette controlled by a synthetic bi-directional promoter, which is composed of human phosphoglycerate kinase (PGK) promoter and minimal core promoter from human cytomegalovirus (mCMV). The expression pattern of DeltaLNGFR and GFP following Ad5F35-DeltaLNGFR-BiDp gene transfer in various cell types including candidate HSCs was compared to Ad5F35-DeltaLNGFR-IRES vector encoding PGK promoter controlled bicistronic expression cassette for DeltaLNGFR and GFP. RESULTS AND CONCLUSIONS: Using Ad5F35-DeltaLNGFR-BiDp, we demonstrated a coordinated, high-level dual gene expression in leukemic cells and cord blood CD34(+) cells. However, the ability of Ad5F35-DeltaLNGFR-BiDp-GFP for coordinated dual gene expression varied significantly between re-populating progenitor cells. In NOD/SCID mice bone marrow transplantation assay, sorted CD34(+)/DeltaLNGFR(+)/GFP(+) cells following infection with Ad5F35-DeltaLNGFR-BiDp showed predominantly myeloid lineage reconstitution with limited lymphoid lineage differentiation capacity, whereas the CD34(+)/DeltaLNGFR(+)/GFP(-) cells exhibited both myeloid and lymphoid reconstitution. This study indicates that bi-directional promoter controlled Ad5F35 vector such as Ad5F35-DeltaLNGFR-BiDp can be particularly useful for manipulation of myeloid progenitor cells and potentially also in myeloid lineage leukemic cells. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1581781
- author
- Na, Manli LU and Fan, Xiaolong LU
- organization
- publishing date
- 2010
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Experimental Hematology
- volume
- Apr 8
- pages
- 446 - 452
- publisher
- Elsevier
- external identifiers
-
- wos:000278178000004
- pmid:20303383
- scopus:77952953570
- ISSN
- 1873-2399
- DOI
- 10.1016/j.exphem.2010.03.007
- language
- English
- LU publication?
- yes
- id
- 5a61e4d6-266f-4455-846c-f739bd1c32d4 (old id 1581781)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/20303383?dopt=Abstract
- date added to LUP
- 2016-04-04 08:41:31
- date last changed
- 2022-04-23 17:53:53
@article{5a61e4d6-266f-4455-846c-f739bd1c32d4,
abstract = {{OBJECTIVE: Adenoviral vector mediated gene expression is an attractive approach to manipulate or report gene expression in human hematopoietic stem cells (HSC), when transient gene expression is preferred. Previous studies have demonstrated that fiber retargeted Ad5F35 vectors can mediate efficient gene transfer into human HSCs. In this study, we have investigated the potential of bi-directional promoter controlled Ad5F35 vector for coordinated dual gene expression in candidate HSCs. MATERIAL AND METHODS: We have engineered Ad5F35-DeltaLNGFR-BiDp encoding kinase domain deleted low-affinity NGF receptor (DeltaLNGFR) and green fluorescent protein (GFP) expression cassette controlled by a synthetic bi-directional promoter, which is composed of human phosphoglycerate kinase (PGK) promoter and minimal core promoter from human cytomegalovirus (mCMV). The expression pattern of DeltaLNGFR and GFP following Ad5F35-DeltaLNGFR-BiDp gene transfer in various cell types including candidate HSCs was compared to Ad5F35-DeltaLNGFR-IRES vector encoding PGK promoter controlled bicistronic expression cassette for DeltaLNGFR and GFP. RESULTS AND CONCLUSIONS: Using Ad5F35-DeltaLNGFR-BiDp, we demonstrated a coordinated, high-level dual gene expression in leukemic cells and cord blood CD34(+) cells. However, the ability of Ad5F35-DeltaLNGFR-BiDp-GFP for coordinated dual gene expression varied significantly between re-populating progenitor cells. In NOD/SCID mice bone marrow transplantation assay, sorted CD34(+)/DeltaLNGFR(+)/GFP(+) cells following infection with Ad5F35-DeltaLNGFR-BiDp showed predominantly myeloid lineage reconstitution with limited lymphoid lineage differentiation capacity, whereas the CD34(+)/DeltaLNGFR(+)/GFP(-) cells exhibited both myeloid and lymphoid reconstitution. This study indicates that bi-directional promoter controlled Ad5F35 vector such as Ad5F35-DeltaLNGFR-BiDp can be particularly useful for manipulation of myeloid progenitor cells and potentially also in myeloid lineage leukemic cells.}},
author = {{Na, Manli and Fan, Xiaolong}},
issn = {{1873-2399}},
language = {{eng}},
pages = {{446--452}},
publisher = {{Elsevier}},
series = {{Experimental Hematology}},
title = {{Design of Ad5F35 vectors for coordinated dual gene expression in candidate human hematopoietic stem cells.}},
url = {{http://dx.doi.org/10.1016/j.exphem.2010.03.007}},
doi = {{10.1016/j.exphem.2010.03.007}},
volume = {{Apr 8}},
year = {{2010}},
}