Abstract
A vast wealth of recent research has seen attempts of using microRNA (miRNA) found in biological fluids in clinical research and medicine. One of the reasons behind this trend is the apparent their high stability of cell-free miRNA conferred by small size and packaging in supramolecular complexes. However, researchers in both basic and clinical settings often face the problem of selecting adequate methods to extract appropriate quality miRNA preparations for use in specific downstream analysis pipelines. This review outlines the variety of different methods of miRNA isolation from biofluids and examines the key determinants of their efficiency, including, but not limited to, the structural properties of miRNA and factors defining their stability in the extracellular environment.
Highlights
The biology of miRNA and their transport from cells and into intracellular space, including biological fluids, were actively studied in the last two decades
Cf-miRNA stability in blood and other biofluids can be attributed to the protection from RNAses conferred by interaction with biomolecules as well as packaging in membrane-coated or membrane-free particles (Table 2). These complexes strongly interfere with the isolation efficacy requiring liberation of miRNA from such structures to ensure effective cf-miRNA isolation and prevent coisolation of polymerase reaction inhibitors
We provide an an2 aolfy1s8is of current methods of cf-miRNA purification from biological fluids and examine factors that affect its efficacy, including intrinsic features of miRNAs, packaging and common contaminants of miRNA preparations
Summary
The biology of miRNA and their transport from cells and into intracellular space, including biological fluids, were actively studied in the last two decades. Cf-miRNA stability in blood and other biofluids can be attributed to the protection from RNAses conferred by interaction with biomolecules as well as packaging in membrane-coated or membrane-free particles (Table 2). These complexes strongly interfere with the isolation efficacy requiring liberation of miRNA from such structures to ensure effective cf-miRNA isolation and prevent coisolation of polymerase reaction inhibitors. We provide an an aolfy1s8is of current methods of cf-miRNA purification from biological fluids and examine factors that affect its efficacy, including intrinsic features of miRNAs, packaging and common contaminants of miRNA preparations. 5-methylcpysateodusedinnooeusri(nimdei-5ntoCe-)(inΨmo)somindoeidfeiicdfiaicttaiintoigonn [8,13]
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