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Sound packing DNA: packing open circular DNA with low-intensity ultrasound.

Park D, Jung BK, Park H, Lee H, Lee G, Park J, Shin U, Won JH, Jo YJ, Chang JW, Lee S, Yoon D, Seo J, Kim CW - Sci Rep (2015)

Bottom Line: Compact packing of DNA is essential to improve the efficiency of gene delivery, which has broad implications in biology and pharmaceutical research.Here we show that low-intensity pulsed ultrasound can pack open circular DNA into supercoil form.We anticipate our results to be a starting point for improved non-viral gene delivery.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Tumor Immunity Medical Research Center, Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.

ABSTRACT
Supercoiling DNA (folding DNA into a more compact molecule) from open circular forms requires significant bending energy. The double helix is coiled into a higher order helix form; thus it occupies a smaller footprint. Compact packing of DNA is essential to improve the efficiency of gene delivery, which has broad implications in biology and pharmaceutical research. Here we show that low-intensity pulsed ultrasound can pack open circular DNA into supercoil form. Plasmid DNA subjected to 5.4 mW/cm(2) intensity ultrasound showed significant (p-values <0.001) supercoiling compared to DNA without exposure to ultrasound. Radiation force induced from ultrasound and dragging force from the fluid are believed to be the main factors that cause supercoiling. This study provides the first evidence to show that low-intensity ultrasound can directly alter DNA topology. We anticipate our results to be a starting point for improved non-viral gene delivery.

No MeSH data available.


Related in: MedlinePlus

The results of electrophoresis in comparison to normal pEGFP, heated pEGFP, and heated pEGFP after ultrasound sonication.lane(1): normal pEGFP vector, lane(2): heated pEGFP vector, lane(3): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 1%, sonication time: 30 s), lane(4): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 10%, sonication time: 30 s).
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f1: The results of electrophoresis in comparison to normal pEGFP, heated pEGFP, and heated pEGFP after ultrasound sonication.lane(1): normal pEGFP vector, lane(2): heated pEGFP vector, lane(3): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 1%, sonication time: 30 s), lane(4): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 10%, sonication time: 30 s).

Mentions: The electrophoretic mobility of normal pEGFP (+), heated pEGFP (No Ultrasound), and heated and sonicated pEGFP were visualized with electrophoresis in Fig. 1. Each topological form of pEGFP is presented as open circular (row A), linear (row B), supercoiled (row C), and circular single-stranded (row D). Most pEGFPs obtained from the purification process are in the supercoiled form and heat changed the overall composition of DNA formation (see Fig. 1 column (1) and (2)). Applying heat led to significant increases in open circular, linear, and circular single-stranded forms of pEGFP (column (2)). After the sonication of low-intensity ultrasound on the heated pEGFPs, the open circular form of pEGFPs decreased significantly and the supercoiled form of pEGFPs increased significantly (column (3) & (4)). This indicated that many of the open circular pEGFPs were packed back to the supercoiled form, though the conformation of repacked pEGFP may not be identical to that of normal supercoiled pEGFP.


Sound packing DNA: packing open circular DNA with low-intensity ultrasound.

Park D, Jung BK, Park H, Lee H, Lee G, Park J, Shin U, Won JH, Jo YJ, Chang JW, Lee S, Yoon D, Seo J, Kim CW - Sci Rep (2015)

The results of electrophoresis in comparison to normal pEGFP, heated pEGFP, and heated pEGFP after ultrasound sonication.lane(1): normal pEGFP vector, lane(2): heated pEGFP vector, lane(3): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 1%, sonication time: 30 s), lane(4): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 10%, sonication time: 30 s).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4402968&req=5

f1: The results of electrophoresis in comparison to normal pEGFP, heated pEGFP, and heated pEGFP after ultrasound sonication.lane(1): normal pEGFP vector, lane(2): heated pEGFP vector, lane(3): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 1%, sonication time: 30 s), lane(4): heated pEGFP vector which is sonicated by ultrasound (pressure: 400 kPa, duty percentage: 10%, sonication time: 30 s).
Mentions: The electrophoretic mobility of normal pEGFP (+), heated pEGFP (No Ultrasound), and heated and sonicated pEGFP were visualized with electrophoresis in Fig. 1. Each topological form of pEGFP is presented as open circular (row A), linear (row B), supercoiled (row C), and circular single-stranded (row D). Most pEGFPs obtained from the purification process are in the supercoiled form and heat changed the overall composition of DNA formation (see Fig. 1 column (1) and (2)). Applying heat led to significant increases in open circular, linear, and circular single-stranded forms of pEGFP (column (2)). After the sonication of low-intensity ultrasound on the heated pEGFPs, the open circular form of pEGFPs decreased significantly and the supercoiled form of pEGFPs increased significantly (column (3) & (4)). This indicated that many of the open circular pEGFPs were packed back to the supercoiled form, though the conformation of repacked pEGFP may not be identical to that of normal supercoiled pEGFP.

Bottom Line: Compact packing of DNA is essential to improve the efficiency of gene delivery, which has broad implications in biology and pharmaceutical research.Here we show that low-intensity pulsed ultrasound can pack open circular DNA into supercoil form.We anticipate our results to be a starting point for improved non-viral gene delivery.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Tumor Immunity Medical Research Center, Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.

ABSTRACT
Supercoiling DNA (folding DNA into a more compact molecule) from open circular forms requires significant bending energy. The double helix is coiled into a higher order helix form; thus it occupies a smaller footprint. Compact packing of DNA is essential to improve the efficiency of gene delivery, which has broad implications in biology and pharmaceutical research. Here we show that low-intensity pulsed ultrasound can pack open circular DNA into supercoil form. Plasmid DNA subjected to 5.4 mW/cm(2) intensity ultrasound showed significant (p-values <0.001) supercoiling compared to DNA without exposure to ultrasound. Radiation force induced from ultrasound and dragging force from the fluid are believed to be the main factors that cause supercoiling. This study provides the first evidence to show that low-intensity ultrasound can directly alter DNA topology. We anticipate our results to be a starting point for improved non-viral gene delivery.

No MeSH data available.


Related in: MedlinePlus