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Dataset of plasmid DNA extraction using different magnetic nanoparticles (MNPs).

ABSTRACT: In this dataset we integrated figures related to bacterial transformation using pBI121 plasmid and complementary analysis for magnetic nanoparticles (MNPs) characterizations. The structural map of pBI121 plasmid was drawn by Vector NTI software using the complete sequence of binary vector pBI121. Escherichia coli bacteria transformed using pBI121 plasmid and were grown on the selection media containing kanamycin. MNPs were characterized by energy dispersive spectroscopy (EDS) and transmission electron microscopy (TEM). Finally, the overall efficiency of different MNPs (Fe3O4, Fe3O4/SiO2, Fe3O4/SiO2/TiO2) in plasmid DNA isolation was compared using gel electrophoresis analysis. The data supplied in this article supports the accompanying publication "Comparative study of three magnetic nano-particles (FeSO4, FeSO4/SiO2, FeSO4/SiO2/TiO2) in plasmid DNA extraction" (H. Rahnama, A. Sattarzadeh, F. Kazemi, N. Ahmadi, F. Sanjarian, Z. Zand, 2016) [1].

SUBMITTER: Rahnama H

PROVIDER: S-EPMC5107585 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Dataset of plasmid DNA extraction using different magnetic nanoparticles (MNPs).

Rahnama H H Sattarzadeh A A Kazemi F F Ahmadi N N Sanjarian F F Zand Z Z

Data in brief 20161026

In this dataset we integrated figures related to bacterial transformation using pBI121 plasmid and complementary analysis for magnetic nanoparticles (MNPs) characterizations. The structural map of pBI121 plasmid was drawn by Vector NTI software using the complete sequence of binary vector pBI121. <i>Escherichia coli</i> bacteria transformed using pBI121 plasmid and were grown on the selection media containing kanamycin. MNPs were characterized by energy dispersive spectroscopy (EDS) and transmis ...[more]

PMID: 27872876

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Project description:Successful systemic gene delivery requires specific tissue targeting as well as efficient intracellular transfection. Increasingly, research laboratories are fabricating libraries of novel nanoparticles, engineering both new biomaterial structures and composition ratios of multicomponent systems. Yet, methods for screening gene delivery vehicles directly in vivo are often low-throughout, limiting the number of candidate nanoparticles that can be investigated. Here, we report a comprehensive, high-throughput method to evaluate a library of polymeric nanoparticles in vivo for tissue-specific gene delivery. The method involves pairing each nanoparticle formulation with a plasmid DNA (pDNA) that harbors a unique nucleotide sequence serving as the identifying "barcode". Using real time quantitative PCR (qPCR) for detection of the barcoded pDNA and quantitative reverse transcription PCR (RT-qPCR) for transcribed barcoded mRNA, we can quantify accumulation and transfection in tissues of interest. The barcode pDNA and primers were designed with sufficient sensitivity and specificity to evaluate multiple nanoparticle formulations per mouse, improving screening efficiency. Using this platform, we evaluated the biodistribution and transfection of 8 intravenously administered poly(beta-amino ester; PBAE) nanoparticle formulations, each with a PBAE polymer of differential structure. Significant levels of nanoparticle accumulation and gene transfection were observed mainly in organs involved in clearance, including spleen, liver, and kidneys. Interestingly, higher levels of transfection of select organs did not necessarily correlate with higher levels of tissue accumulation, highlighting the importance of directly measuring in vivo transfection efficiency as the key barcoded parameter in gene delivery vector optimization. To validate this method, nanoparticle formulations were used individually for luciferase pDNA delivery in vivo. The distribution of luciferase expression in tissues matched the transfection analysis by the barcode qPCR method, confirming that this platform can be used to accurately evaluate systemic gene delivery.

Dataset Information

Dataset of plasmid DNA extraction using different magnetic nanoparticles (MNPs).

Publications

Dataset of plasmid DNA extraction using different magnetic nanoparticles (MNPs).

Similar Datasets

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