Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro

Current viral gene delivery vectors for gene therapy are inefficient due to short-lived transgene expression attributed to the cytosine-phosphate-guanine (CpG) motifs in the transgene. Here we assessed the effects of CpG motif reduction in lentiviral (LV) gene delivery context on the level and durat...

Full description

Bibliographic Details
Main Authors: Low, Poh Tee, Lai, Mei I., Ngai, Siew Ching, Abdullah, Syahrilnizam
Format: Article
Published: Elsevier 2014
Online Access:http://psasir.upm.edu.my/id/eprint/35891/
_version_ 1848848181033435136
author Low, Poh Tee
Lai, Mei I.
Ngai, Siew Ching
Abdullah, Syahrilnizam
author_facet Low, Poh Tee
Lai, Mei I.
Ngai, Siew Ching
Abdullah, Syahrilnizam
author_sort Low, Poh Tee
building UPM Institutional Repository
collection Online Access
description Current viral gene delivery vectors for gene therapy are inefficient due to short-lived transgene expression attributed to the cytosine-phosphate-guanine (CpG) motifs in the transgene. Here we assessed the effects of CpG motif reduction in lentiviral (LV) gene delivery context on the level and duration of reporter gene expression in Chinese Hamster Ovary (CHO) cells, Human Immortalized Myelogenous Leukemia (K562) cells and hematopoietic stem cells (HSCs). The cells were transduced with LV carrying Zero-CpG green fluorescent protein (ZGFP) reporter gene, LV/CMV/ZGFP. The GFP expression was compared to its non CpG-depleted GFP reporter gene LV (LV/CMV/GFP) counterpart. The LV/CMV/ZGFP exhibited prolonged transgene expression in CHO cells and HSCs up to 10 days and 14 days, in the respective cells. This effect was not seen in the transduced K562 cells, which may be due to the DNA hypomethylation status of the cancer cell line. Transgene copy number analysis verified that the GFP expression was not from pseudo-transduction and the transgene remained in the genome of the cells throughout the period of the study. The modest positive effects from the LV/CMV/ZGFP suggest that the reduction of CpG in the LV construct was not substantial to generate higher and more prolonged transgene expression.
first_indexed 2025-11-15T09:30:25Z
format Article
id upm-35891
institution Universiti Putra Malaysia
institution_category Local University
last_indexed 2025-11-15T09:30:25Z
publishDate 2014
publisher Elsevier
recordtype eprints
repository_type Digital Repository
spelling upm-358912016-02-11T05:04:41Z http://psasir.upm.edu.my/id/eprint/35891/ Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro Low, Poh Tee Lai, Mei I. Ngai, Siew Ching Abdullah, Syahrilnizam Current viral gene delivery vectors for gene therapy are inefficient due to short-lived transgene expression attributed to the cytosine-phosphate-guanine (CpG) motifs in the transgene. Here we assessed the effects of CpG motif reduction in lentiviral (LV) gene delivery context on the level and duration of reporter gene expression in Chinese Hamster Ovary (CHO) cells, Human Immortalized Myelogenous Leukemia (K562) cells and hematopoietic stem cells (HSCs). The cells were transduced with LV carrying Zero-CpG green fluorescent protein (ZGFP) reporter gene, LV/CMV/ZGFP. The GFP expression was compared to its non CpG-depleted GFP reporter gene LV (LV/CMV/GFP) counterpart. The LV/CMV/ZGFP exhibited prolonged transgene expression in CHO cells and HSCs up to 10 days and 14 days, in the respective cells. This effect was not seen in the transduced K562 cells, which may be due to the DNA hypomethylation status of the cancer cell line. Transgene copy number analysis verified that the GFP expression was not from pseudo-transduction and the transgene remained in the genome of the cells throughout the period of the study. The modest positive effects from the LV/CMV/ZGFP suggest that the reduction of CpG in the LV construct was not substantial to generate higher and more prolonged transgene expression. Elsevier 2014-01-01 Article PeerReviewed Low, Poh Tee and Lai, Mei I. and Ngai, Siew Ching and Abdullah, Syahrilnizam (2014) Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro. Gene, 533 (1). pp. 451-455. ISSN 0378-1119; ESSN: 1879-0038 http://www.sciencedirect.com/science/article/pii/S037811191301295X 10.1016/j.gene.2013.09.075
spellingShingle Low, Poh Tee
Lai, Mei I.
Ngai, Siew Ching
Abdullah, Syahrilnizam
Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro
title Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro
title_full Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro
title_fullStr Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro
title_full_unstemmed Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro
title_short Transgene expression from CpG-reduced lentiviral gene delivery vectors in vitro
title_sort transgene expression from cpg-reduced lentiviral gene delivery vectors in vitro
url http://psasir.upm.edu.my/id/eprint/35891/
http://psasir.upm.edu.my/id/eprint/35891/
http://psasir.upm.edu.my/id/eprint/35891/