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Transient Gene Expression Following Single and Double Exponential Decay Electroporation Pulses

ase or CD2 expression several fold. The electroporation conditions were optimized for voltage, but not for capacitance, resistance, or timing between pulses. Furthermore, these researchers did not compare the results of single, exponential decay pulses to the expression produced by the double pulse procedure. De Chasseval and de Villartay did not claim that the use of the double pulse was essential for the success of these experiments.

In this report, the optimal Twin-pulse electroporation conditions from de Chasseval and de Villarty, as reported in the January, 1993 EquiBio News, were compared to single electroporation pulses generated by the Gene Pulser II system for transient gene expression in COS cells.


Results and Discussion
The results in Table 1 and Figure 1 show the reporter gene activity obtained for optimal electroporation conditions of the two instruments. Maximum transient expression was obtained with both the Gene Pulser II system and the EasyJect Plus with a single pulse of 210V and 450 F capacitance. The third treatment duplicates the optimum Twin-pulse conditions reported by EquiBio for mammalian cells a 500 V, 25 F pulse with a 99 ohm resistor in parallel, a 7 second inter-pulse delay, and then a 74 V, 2,100 F pulse with a 99 ohm resistor in parallel.

It is clearly apparent that a single, optimized exponential decay pulse is more effective than this Twin-pulse form, generating about twice the level of transient gene expression. Moreover, when the two parts of the double pulse were delivered to cell samples separately in treatments 4 and 5, ca. 70% of the Twinpulse expression was obtained when the first, high voltage component
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