RNA extraction from self-assembling peptide hydrogels to allow qPCR analysis

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dc.cclicenceCC-BY-NCen
dc.contributor.authorBurgess, K.
dc.contributor.authorWorkman, V.
dc.contributor.authorElsawy, M.
dc.contributor.authorMiller, A.
dc.contributor.authorOceandy, D.
dc.contributor.authorSaiani, A.
dc.date.accessioned2020-01-28T14:47:08Z
dc.date.available2020-01-28T14:47:08Z
dc.date.issued2018-06-04
dc.descriptionopen access articleen
dc.description.abstractSelf-assembling peptide hydrogels offer a novel 3-dimensional platform for many applications in cell culture and tissue engineering but are not compatible with current methods of RNA isolation; owing to interactions between RNA and the biomaterial. This study investigates the use of two techniques based on two different basic extraction principles: solution-based extraction and direct solid-state binding of RNA respectively, to extract RNA from cells encapsulated in four β-sheet forming self-assembling peptide hydrogels with varying net positive charge. RNA-peptide fibril interactions, rather than RNA-peptide molecular complexing, were found to interfere with the extraction process resulting in low yields. A column-based approach relying on RNA-specific binding was shown to be more suited to extracting RNA with higher purity from these peptide hydrogels owing to its reliance on strong specific RNA binding interactions which compete directly with RNA-peptide fibril interactions. In order to reduce the amount of fibrils present and improve RNA yields a broad spectrum enzyme solution—pronase—was used to partially digest the hydrogels before RNA extraction. This pre-treatment was shown to significantly increase the yield of RNA extracted, allowing downstream RT-qPCR to be performed.en
dc.exception.ref2021codes254aen
dc.funderEPSRC (Engineering and Physical Sciences Research Council)en
dc.identifier.citationBurgess, K., Workman, V., Elsawy, M., Miller, A., Oceandy, D., Saiani, A. (2018) RNA extraction from self-assembling peptide hydrogels to allow qPCR analysis. PLOS ONE, 13, e0197517.en
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0197517
dc.identifier.urihttps://dora.dmu.ac.uk/handle/2086/19081
dc.language.isoenen
dc.peerreviewedYesen
dc.publisherPLOSen
dc.researchinstituteLeicester Institute for Pharmaceutical Innovation - From Molecules to Practice (LIPI)en
dc.subjectNanomaterialsen
dc.subjectHydrogelsen
dc.subjectPeptidesen
dc.subjectSelf-assemblyen
dc.subjectFibersen
dc.subjectRNA extractionen
dc.subjectMolecular Biologyen
dc.subject3D Cell Cultureen
dc.titleRNA extraction from self-assembling peptide hydrogels to allow qPCR analysisen
dc.typeArticleen

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