Stable gene transfer to human CD34 + hematopoietic cells using the Sleeping Beauty transposon

Methods of gene transfer to hematopoietic stem cells that result in stable integration may provide treatments for many inherited and acquired blood diseases. It has been demonstrated previously that a gene delivery system based on the Sleeping Beauty (SB) transposon can be derived where a plasmid tr...

Full description

Saved in:
Bibliographic Details
Published in:Experimental hematology 2006-10, Vol.34 (10), p.1333-1343
Main Authors: Hollis, Roger P., Nightingale, Sarah J., Wang, Xiuli, Pepper, Karen A., Yu, Xiao-Jin, Barsky, Lora, Crooks, Gay M., Kohn, Donald B.
Format: Article
Language:English
Subjects:
Antigens, CD34
DNA Transposable Elements - genetics
Electroporation - methods
Gene Expression
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - metabolism
Humans
K562 Cells
Mutagenesis, Insertional - methods
Time Factors
Transposases - biosynthesis
Transposases - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Methods of gene transfer to hematopoietic stem cells that result in stable integration may provide treatments for many inherited and acquired blood diseases. It has been demonstrated previously that a gene delivery system based on the Sleeping Beauty (SB) transposon can be derived where a plasmid transiently expressing the SB transposase can mediate the stable chromosomal integration of a codelivered second plasmid containing a gene expression unit flanked by the inverted repeats derived from the transposon. Plasmid DNA containing the elements required for SB transposition was delivered to hematopoietic cells via electroporation. Integrated transgene (enhanced green fluorescent protein [eGFP]) expression was assessed in vitro and in vivo. In the K562 human hematopoietic cell line, we observed stable expression of eGFP in >60% of cells for over 2 months after electroporation of the two plasmids; in contrast, in control cells either not treated with transposase or exposed to a defective mutant transposase, the level of gene expression had fallen to near background (
ISSN:0301-472X
1873-2399
DOI:10.1016/j.exphem.2006.05.023