ArticleNucleic acids research2010
A rapid and sensitive method to detect siRNA-mediated mRNA cleavage in vivo using 5' RACE and a molecular beacon probe.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 24 citations in OpenAlex.
- N-Methyl-D-Aspartate Receptor Hypofunction in Meg-01 Cells Reveals a Role for Intracellular Calcium Homeostasis in Balancing Megakaryocytic-Erythroid Differentiation.Thrombosis and haemostasis · 2020Article
- Preassembled Single-Stranded RNA-Argonaute Complexes: A Novel Method to Silence Genes in Cryptosporidium.The Journal of infectious diseases · 2016Article
- Antibody targeting facilitates effective intratumoral siRNA nanoparticle delivery to HER2-overexpressing cancer cells.Oncotarget · 2016Article
- Quantitative analysis of deadenylation-independent mRNA decay by a modified MBRACE assay.Methods in molecular biology (Clifton, N.J.) · 2014Article
- SMG6 cleavage generates metastable decay intermediates from nonsense-containing β-globin mRNA.PloS one · 2013Article
- Article
- Multispecies-compatible antitumor effects of a cross-species small-interfering RNA against mammalian target of rapamycin.Cellular and molecular life sciences : CMLS · 2012Article
- Silencing disease genes in the laboratory and the clinic.The Journal of pathology · 2012Review
- Anti-CD22 antibody targeting of pH-responsive micelles enhances small interfering RNA delivery and gene silencing in lymphoma cells.Molecular therapy : the journal of the American Society of Gene Therapy · 2011Article
- The importance of RT-qPCR primer design for the detection of siRNA-mediated mRNA silencing.BMC research notes · 2011Article
- Gene regulation by small RNAs.Journal of RNAi and gene silencing : an international journal of RNA and gene targeting research · 2010Article
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Specific detection of mRNA cleavage by 5'RACE is the only method to confirm the knockdown of mRNA by RNA interference, but is rarely reported for in vivo studies. We have combined 5'-RNA-linker-mediated RACE (5'-RLM-RACE) with real-time PCR using a molecular beacon to develop a rapid and specific method termed MBRACE, which we have used to detect small-interfering RNA (siRNA)-induced cleavage of ApoB, RRM1 and YBX1 transcripts in vitro, and ApoB in vivo. When RNA from siRNA-transfected cells was used for 5'-RLM-RACE and a cleavage site-specific molecular beacon probe was included in subsequent real-time PCR analysis, the specific mRNA cleavage product was detected. Detection of siRNA-mediated cleavage was also observed when RNA from mouse liver following administration of ApoB-specific siRNA was analysed, even in cases where ApoB knockdown measured by real-time PCR was <10%. With its sensitivity and specificity, this variation on the 5'RACE method should prove a useful tool to detect mRNA cleavage and corroborate knockdown studies following siRNA use in vivo.
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