NDLI: DNA amplification via polymerase chain reaction inside miniemulsion droplets with subsequent poly(n-butylcyanoacrylate) shell formation and delivery of polymeric capsules into mammalian cells.

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DNA amplification via polymerase chain reaction inside miniemulsion droplets with subsequent poly(n-butylcyanoacrylate) shell formation and delivery of polymeric capsules into mammalian cells.

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DNA amplification via polymerase chain reaction inside miniemulsion droplets with subsequent poly(n-butylcyanoacrylate) shell formation and delivery of polymeric capsules into mammalian cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Baier, Grit Musyanovych, Anna Landfester, Katharina Best, Andreas Lorenz, Steffen Mailänder, Volker
Description	Country affiliation: Germany Author Affiliation: Baier G ( Max Planck Institute for Polymer Research, Ackermannweg 10, Mainz, Germany.)
Abstract	There is a growing interest in the development of stable nanocapsules that could deliver the bioactive compounds within the living organism, and to release them without causing any toxic effects. Here the miniemulsion droplets were first used as 'nanoreactors' for the amplification of single-molecule dsDNA template (476 and 790 base pairs) through PCR. Afterwards, each droplet was surrounded with a biodegradable PBCA shell by interfacial anionic polymerization, enabling therefore to deliver the PCR products into the cells. The size of the initial miniemulsion droplets and the final polymeric capsules was in the range of 250 and 320 nm, mainly depending on the type of the continuous phase and presence of dsDNA template molecules. The formation of PCR products was resolved with gel electrophoresis and detected with fluorescence spectroscopy in the presence of DNA specific dye (SYBRGreen). TEM studies were performed to prove the formation of the polymeric shell. The shell thickness was measured to be within 5-15 nm and the average molecular weight of the formed PBCA polymer was around 75000 g · mol(-1) . For the cell uptake experiments, the obtained nanocapsules were transferred from the organic phase into aqueous medium containing a water-soluble surfactant. The effect of the surfactant type (anionic, cationic or non-ionic) on the HeLa cell viability and nanocapsule uptake behavior was studied by CLSM and FACS. Confocal analysis demonstrated that nanocapsules stabilized with cationic (CTMA-Cl) and non-ionic (Lutensol AT50) surfactants show almost the same uptake, whereas capsules redispersed in anionic (SDS) surfactant possess a 30% higher uptake. The release of the encapsulated material within the cell was studied on the example of Cy5-labeled oligonucleotides showing the colocalization with mitochondria of MSCs cells.
File Format	HTM / HTML
ISSN	16165187
Issue Number	8
Volume Number	11
e-ISSN	16165195
Journal	Macromolecular Bioscience
Language	English
Publisher	Wiley-VCH
Publisher Date	2011-08-11
Publisher Place	Germany
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Biochemistry Dna Metabolism Drug Delivery Systems Methods Emulsions Chemistry Enbucrilate Chemical Synthesis Nanocapsules Nanotechnology Nucleic Acid Amplification Techniques Carbocyanines Analysis Pharmacology Electrophoresis, Polyacrylamide Gel Female Flow Cytometry Hela Cells Humans Microscopy, Fluorescence Mitochondria Drug Effects Molecular Weight Oils Polymerase Chain Reaction Surface-active Agents Triglycerides Journal Article
Content Type	Text
Resource Type	Article
Subject	Materials Chemistry Biomaterials Bioengineering Biotechnology Polymers and Plastics

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DNA amplification via polymerase chain reaction inside miniemulsion droplets with subsequent poly(n-butylcyanoacrylate) shell formation and delivery of polymeric capsules into mammalian cells.

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