caa a22 4500 477125794 CHVBK 20180405111649.0 cr unu---uuuuu 170330e19970601xx s 000 0 eng 10.1023/A:1012198321879 doi (NATIONALLICENCE)springer-10.1023/A:1012198321879 The Use of Sonication for the Efficient Delivery of Plasmid DNA into Cells [Elektronische Daten] [Julie Wyber, Julie Andrews, Antony D'Emanuele] Purpose. Ultrasonic methods have considerable potential for the introduction of macromolecules into cells. In this paper we demonstrate that, under controlled conditions, application of 20kHz ultrasound to a suspension of yeast cells facilitates the delivery of plasmid DNA into these cells. Methods. Aliquots of growing yeast cells (Saccharomyces cerevisae, strain AH22) were suspended in buffer and exposed to 20kHz ultrasound from a laboratory (probe-type) sonicator in the presence of microgram quantities of plasmid DNA. Efficiency of DNA delivery was scored as the number of cells transformed. Results. Cell transformation was optimal at 30 seconds sonication using an output of 2.0 watts and resulted in a 20 fold enhancement over control values. At extended sonication times, fewer cells showed evidence of transformation because of reduced cell viability. The increased DNA uptake and the decreased cell viability were both attributable to acoustic cavitation events during sonication. The extent of acoustic cavitation was measured and it was found that there was an increase in cavitation events with increased sonication time. Cell viability was shown to be directly related to the number of cavitation events. The effects of sonication on plasmid DNA were investigated and indicated that the structural integrity of plasmid DNA was unaffected by the sonication conditions employed. Conclusions. Under controlled conditions, ultrasound is an effective means of delivering plasmid DNA into cells. The subsequent expression of DNA molecules in cells depends upon a balance between transient cell damage and cell death. Plenum Publishing Corporation, 1997 DNA delivery nationallicence cells nationallicence sonication nationallicence cavitation nationallicence transformation nationallicence Wyber Julie School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Ml3 9PL, Manchester, UK aut Andrews Julie School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Ml3 9PL, Manchester, UK aut D'Emanuele Antony School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Ml3 9PL, Manchester, UK aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/6(1997-06-01), 750-756 0724-8741 14:6<750 1997 14 11095 https://doi.org/10.1023/A:1012198321879 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012198321879 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wyber Julie School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Ml3 9PL, Manchester, UK aut NATIONALLICENCE 700 1- Andrews Julie School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Ml3 9PL, Manchester, UK aut NATIONALLICENCE 700 1- D'Emanuele Antony School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Ml3 9PL, Manchester, UK aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/6(1997-06-01), 750-756 0724-8741 14:6<750 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer