Sleeping beauty-mediated drug resistance gene transfer in human hematopoietic progenitor cells

Kendra A. Hyland, Erik R. Olson, R. Scott Mcivor

Research output: Contribution to journalArticle

Abstract

The Sleeping Beauty (SB) transposon system can insert sequences into mammalian chromosomes, supporting long-term expression of both reporter and therapeutic genes. Hematopoietic progenitor cells (HPCs) are an ideal therapeutic gene transfer target as they are used in therapy for a variety of hematologic and metabolic conditions. As successful SB-mediated gene transfer into human CD34+ HPCs has been reported by several laboratories, we sought to extend these studies to the introduction of a therapeutic gene conferring resistance to methotrexate (MTX), potentially providing a chemoprotective effect after engraftment. SB-mediated transposition of hematopoietic progenitors, using a transposon encoding an L22Y variant dihydrofolate reductase fused to green fluorescent protein, conferred resistance to methotrexate and dipyridamole, a nucleoside transport inhibitor that tightens MTX selection conditions, as assessed by in vitro hematopoietic colony formation. Transposition of individual transgenes was confirmed by sequence analysis of transposon-chromosome junctions recovered by linear amplification-mediated PCR. These studies demonstrate the potential of SB-mediated transposition of HPCs for expression of drug resistance genes for selective and chemoprotective applications.

Original languageEnglish (US)
Pages (from-to)657-663
Number of pages7
JournalHuman gene therapy
Volume26
Issue number10
DOIs
StatePublished - Oct 1 2015

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Beauty
Hematopoietic Stem Cells
Drug Resistance
Methotrexate
Genes
Mammalian Chromosomes
Tetrahydrofolate Dehydrogenase
Dipyridamole
Therapeutics
Green Fluorescent Proteins
Transgenes
Reporter Genes
Nucleosides
Sequence Analysis
Chromosomes
Polymerase Chain Reaction

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Sleeping beauty-mediated drug resistance gene transfer in human hematopoietic progenitor cells. / Hyland, Kendra A.; Olson, Erik R.; Mcivor, R. Scott.

In: Human gene therapy, Vol. 26, No. 10, 01.10.2015, p. 657-663.

Research output: Contribution to journalArticle

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