Intragenic integration in DLC1 sustains factor VIII expression in primary human cells without insertional oncogenicity

Techniques enabling precise genome modifications enhance the safety of gene-based therapy. DLC1 is a hot spot for phiC31 integrase-mediated transgene integration in vitro and in vivo . Here we show that integration of a coagulation factor VIII transgene into intron 7 of DLC1 supports durable express...

Full description

Saved in:
Bibliographic Details
Published inGene therapy Vol. 21; no. 4; pp. 402 - 412
Main Authors Sivalingam, J, Phan, T T, Kon, O L
Format Journal Article
LanguageEnglish
Published London Nature Publishing Group UK 01.04.2014
Nature Publishing Group
Subjects
631/61/201
631/61/51/1844
Base Sequence
Biomedical and Life Sciences
Biomedicine
Blood coagulation factor VIII
Cell Biology
Cells
Deoxyribonucleic acid
DNA
DNA Copy Number Variations
Factor VIII - biosynthesis
Gene Expression
Gene Expression Regulation, Neoplastic
Gene Therapy
Genetic engineering
Genetic research
Genetic Therapy
GTPase-Activating Proteins - genetics
Human Genetics
Human Umbilical Vein Endothelial Cells
Humans
Integrases - genetics
Mutagenesis, Insertional
Nanotechnology
Original
original-article
Properties
Transfection
Transgenes
Transgenic plants
Tumor Suppressor Proteins - genetics
Online AccessGet full text

Cover

More Information
Summary:Techniques enabling precise genome modifications enhance the safety of gene-based therapy. DLC1 is a hot spot for phiC31 integrase-mediated transgene integration in vitro and in vivo . Here we show that integration of a coagulation factor VIII transgene into intron 7 of DLC1 supports durable expression of factor VIII in primary human umbilical cord-lining epithelial cells. Oligoclonal cells with factor VIII transgene integrated in DLC1 did not have altered expression of DLC1 or neighbouring genes within a 1-Mb interval. Only 1.9% of all expressed genes were transcriptionally altered; most were downregulated and mapped to cell cycle and DNA repair pathways. DLC1 -integrated cells were not tumourigenic in vivo and were normal by high-resolution genomic DNA copy number analysis. Our data identify DLC1 as a locus for durable transgene expression that does not incur features of insertional oncogenesis, thus expanding options for developing ex vivo cell therapy mediated by site-specific integration methods.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ISSN:0969-7128
1476-5462
DOI:10.1038/gt.2014.11