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Human Gene Therapy
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Calcium Electrotransfer for Termination of Transgene Expression in Muscle

Authors: Hojman, Pernille; Spanggaard, Iben; Olsen, Caroline Holkman; Gehl, Julie; Gissel, Hanne;

Calcium Electrotransfer for Termination of Transgene Expression in Muscle

Abstract

Gene electrotransfer is expanding in clinical use, thus we have searched for an emergency procedure to stop transgene expression in case of serious adverse events. Calcium is cytotoxic at high intracellular levels, so we tested effects of calcium electrotransfer on transgene expression in muscle. A clinical grade calcium solution (20 μl, 168 m M ) was injected into transfected mouse or rat tibialis cranialis muscle. Ca 2+ uptake was quantified using calcium 45 ( 45 Ca), and voltage and time between injection and pulsation were varied. Extinction of transgene expression was investigated by using both in vivo imaging of infrared fluorescent “Katushka” and erythropoietin evaluated by ELISA and hemoglobin. Histology was performed. Electrotransfer of Katushka and erythropoietin yielded significant expression. Maximal calcium uptake occurred after injection of Ca 2+ before electropulsing using eight high voltage pulses of 1000 V/cm. Using these parameters, in vivo imaging showed that transgene expression significantly decreased 4 hr after Ca 2+ electrotransfer and was eliminated within 24 hr. Similarly, serum erythropoietin was reduced by 46% at 4 hr and to control levels at 2 days. Histological analyses showed muscle damage and subsequent regeneration. Electrotransfer of isotonic CaCl 2 terminates transgenic protein expression in muscles and may be used for contingency elimination of transgene expression.

Country
Denmark
Keywords

Muscles, Gene Transfer Techniques, Gene Expression, Gene Therapy, Genetic Therapy, Rats, Mice, Inbred C57BL, Mice, Electroporation, Animals, Calcium, Transgenes, Rats, Wistar, Erythropoietin

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    influence
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    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
17
Top 10%
Top 10%
Top 10%
bronze