FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W4246097986> ?p ?o ?g. }

• W4246097986 endingPage "60" @default.
• W4246097986 startingPage "60" @default.
• W4246097986 abstract "<ns4:p>RNA interference (RNAi) is widely used to suppress gene expression in a specific manner. The efficacy of RNAi is mainly dependent on the sequence of small interfering RNA (siRNA) in relation to the target mRNA. Although several algorithms have been developed for the design of siRNA, it is still difficult to choose a really effective siRNA from among multiple candidates. In this article, we report the development of an image-based, quantitative, ratiometric fluorescence reporter assay to evaluate the efficacy of RNAi at the single-cell level. Two fluorescence reporter constructs are used. One expresses the candidate small hairpin RNA (shRNA) together with an enhanced green fluorescent protein (EGFP); the other expresses a 19-nt target sequence inserted into a cassette expressing a red fluorescent protein (either DsRed or mCherry). Effectiveness of the candidate shRNA is evaluated as the extent to which it knocks down expression of the red fluorescent protein. Thus, the red-to-green fluorescence intensity ratio (appropriately normalized to controls) is used as the read-out for quantifying the siRNA efficacy at the individual cell level. We tested this dual fluorescence assay and compared predictions to actual endogenous knockdown levels for three different genes (vimentin, lamin A/C and Arp3) and twenty different shRNAs. For each of the genes, our assay successfully predicted the target sequences for effective RNAi.</ns4:p><ns4:p>To further facilitate testing of RNAi efficacy, we developed a negative selection marker (<ns4:italic>ccdB</ns4:italic>) method for construction of shRNA and red fluorescent reporter plasmids that allowed us to purify these plasmids directly from transformed bacteria without the need for colony selection and DNA sequencing verification.</ns4:p>" @default.
• W4246097986 created "2022-05-12" @default.
• W4246097986 creator A5002316771 @default.
• W4246097986 creator A5082853086 @default.
• W4246097986 date "2014-05-20" @default.
• W4246097986 modified "2023-09-28" @default.
• W4246097986 title "An image-based, dual fluorescence reporter assay to evaluate the efficacy of shRNA for gene silencing at the single-cell level" @default.
• W4246097986 cites W1611419331 @default.
• W4246097986 cites W1647075334 @default.
• W4246097986 cites W1972748508 @default.
• W4246097986 cites W1977943896 @default.
• W4246097986 cites W1978129709 @default.
• W4246097986 cites W2003171404 @default.
• W4246097986 cites W2019387053 @default.
• W4246097986 cites W2025074364 @default.
• W4246097986 cites W2026520402 @default.
• W4246097986 cites W2030621748 @default.
• W4246097986 cites W2036176224 @default.
• W4246097986 cites W2056022845 @default.
• W4246097986 cites W2064815984 @default.
• W4246097986 cites W2071142904 @default.
• W4246097986 cites W2077659966 @default.
• W4246097986 cites W2079707100 @default.
• W4246097986 cites W2098100188 @default.
• W4246097986 cites W2098270492 @default.
• W4246097986 cites W2112444085 @default.
• W4246097986 cites W2115734610 @default.
• W4246097986 cites W2137315132 @default.
• W4246097986 cites W2138841516 @default.
• W4246097986 cites W2145638174 @default.
• W4246097986 cites W2149402050 @default.
• W4246097986 cites W2149538815 @default.
• W4246097986 cites W2152993806 @default.
• W4246097986 cites W2162674813 @default.
• W4246097986 cites W2166802510 @default.
• W4246097986 cites W2170781554 @default.
• W4246097986 cites W2249233795 @default.
• W4246097986 cites W80812699 @default.
• W4246097986 doi "https://doi.org/10.12688/f1000research.3-60.v2" @default.
• W4246097986 hasPublicationYear "2014" @default.
• W4246097986 type Work @default.
• W4246097986 citedByCount "3" @default.
• W4246097986 countsByYear W42460979862016 @default.
• W4246097986 countsByYear W42460979862021 @default.
• W4246097986 crossrefType "journal-article" @default.
• W4246097986 hasAuthorship W4246097986A5002316771 @default.
• W4246097986 hasAuthorship W4246097986A5082853086 @default.
• W4246097986 hasBestOaLocation W42460979861 @default.
• W4246097986 hasConcept C104317684 @default.
• W4246097986 hasConcept C119056186 @default.
• W4246097986 hasConcept C142613039 @default.
• W4246097986 hasConcept C150194340 @default.
• W4246097986 hasConcept C153911025 @default.
• W4246097986 hasConcept C161733203 @default.
• W4246097986 hasConcept C166703698 @default.
• W4246097986 hasConcept C173396325 @default.
• W4246097986 hasConcept C190232843 @default.
• W4246097986 hasConcept C22615655 @default.
• W4246097986 hasConcept C2779260011 @default.
• W4246097986 hasConcept C54355233 @default.
• W4246097986 hasConcept C67705224 @default.
• W4246097986 hasConcept C86803240 @default.
• W4246097986 hasConcept C95444343 @default.
• W4246097986 hasConceptScore W4246097986C104317684 @default.
• W4246097986 hasConceptScore W4246097986C119056186 @default.
• W4246097986 hasConceptScore W4246097986C142613039 @default.
• W4246097986 hasConceptScore W4246097986C150194340 @default.
• W4246097986 hasConceptScore W4246097986C153911025 @default.
• W4246097986 hasConceptScore W4246097986C161733203 @default.
• W4246097986 hasConceptScore W4246097986C166703698 @default.
• W4246097986 hasConceptScore W4246097986C173396325 @default.
• W4246097986 hasConceptScore W4246097986C190232843 @default.
• W4246097986 hasConceptScore W4246097986C22615655 @default.
• W4246097986 hasConceptScore W4246097986C2779260011 @default.
• W4246097986 hasConceptScore W4246097986C54355233 @default.
• W4246097986 hasConceptScore W4246097986C67705224 @default.
• W4246097986 hasConceptScore W4246097986C86803240 @default.
• W4246097986 hasConceptScore W4246097986C95444343 @default.
• W4246097986 hasLocation W42460979861 @default.
• W4246097986 hasLocation W42460979862 @default.
• W4246097986 hasOpenAccess W4246097986 @default.
• W4246097986 hasPrimaryLocation W42460979861 @default.
• W4246097986 hasRelatedWork W2075512174 @default.
• W4246097986 hasRelatedWork W2088835981 @default.
• W4246097986 hasRelatedWork W2093704434 @default.
• W4246097986 hasRelatedWork W2139684752 @default.
• W4246097986 hasRelatedWork W2150505140 @default.
• W4246097986 hasRelatedWork W2373539659 @default.
• W4246097986 hasRelatedWork W2413319726 @default.
• W4246097986 hasRelatedWork W2414655079 @default.
• W4246097986 hasRelatedWork W4246097986 @default.
• W4246097986 hasRelatedWork W80812699 @default.
• W4246097986 hasVolume "3" @default.
• W4246097986 isParatext "false" @default.
• W4246097986 isRetracted "false" @default.
• W4246097986 workType "article" @default.