Pre-microrna Profiling
Pre-microrna Profiling - Related Videos
10:58
Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays
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Cited by 17 •
2010
We will demonstrate the setup and analysis of pre-microRNA 96-well arrays for QPCR using a robot as well as by hand with a Thermo Scientific Matrix multichannel pipette.
06:59
Isolation and Profiling of MicroRNA-containing Exosomes from Human Bile
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Cited by 17 •
2016
Bile fluid is a valuable source of extracellular vesicles/exosomes that contain potentially important biomarkers. This protocol represents a robust method to isolate exosomes from human bile for further analyses including miRNA profiling.
09:26
Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR
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Cited by 10 •
2014
We describe a protocol of real time PCR to profile microRNAs in the cerebrospinal fluid (CSF). With the exception of RNA extraction protocols, the procedure can be extended to RNA extracted from other body fluids, cultured cells, or tissue specimens.
16:24
Profiling of Estrogen-regulated MicroRNAs in Breast Cancer Cells
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Cited by 9 •
2014
Molecular signaling through both estrogen and microRNAs are critical in breast cancer development and growth. Estrogen activates the estrogen receptors, which are transcription factors. Many transcription factors can regulate the expression of microRNAs, and estrogen-regulated microRNAs can be profiled using different large-scale techniques.
10:45
Purification and microRNA Profiling of Exosomes Derived from Blood and Culture Media
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Cited by 45 •
2013
The presence of stable microRNAs (miRNAs) in exosomes has generated immense interest as a novel mode of intercellular communication, for their potential utility as biomarkers and as a route for therapeutic intervention. Here we demonstrate exosome purification from blood and culture media followed by quantitative PCR to identify miRNAs being transported.
FAQs
The precursor’s position in the canonical pathway determines what a profile can reveal. Drosha-DGCR8 cleavage occurs in the nucleus, Exportin-5 moves the hairpin to the cytoplasm, and Dicer performs a later cut there. Measuring pre-miRNA therefore provides stage-specific information that can indicate whether altered regulation is associated with processing before or after nuclear export.
Comparing both forms separates changes in precursor production from changes in downstream processing or mature-miRNA accumulation. A tissue or condition may show altered pre-miRNA levels without a matching mature-miRNA change, or the reverse. This comparison helps investigators determine whether regulation is occurring during biogenesis or at a later stage of gene-expression control.
Pre-miRNA patterns can vary among tissues and across biological conditions because the activity of the miRNA biogenesis pathway is not necessarily uniform. Profiling these differences helps identify condition-specific regulation rather than treating miRNA production as constant. In biology, such comparisons can connect precursor changes with development, disease, or cellular responses.
RNA sequencing and targeted reverse-transcription assays provide complementary ways to detect and quantify precursor molecules. Sequencing supports characterization across a broader set of RNA species, whereas a targeted assay focuses measurement on selected pre-miRNAs. The choice depends on whether the study emphasizes wider profiling or focused examination of particular candidate regulators.
A basic workflow selects either RNA sequencing or a targeted reverse-transcription assay, measures precursor abundance, and compares the resulting profile across tissues or experimental conditions. Researchers then interpret those measurements alongside mature-miRNA levels and the precursor’s position in the processing pathway. This approach links observed abundance changes to possible differences in miRNA biogenesis.
The measurements can reveal tissue- and condition-specific patterns and help identify candidate regulators involved in development, disease, and cellular responses. Because precursor abundance reflects an intermediate stage, the data can also distinguish altered biogenesis from a change observed only among mature miRNAs. These outcomes make profiling useful for connecting pathway behavior with biological states.