Cloning and expression profiling of testis-expressed microRNAs

摘要

UsinganewsmallRNAcloningmethod,weidentified141miRNAsfromthemousetestis,ofwhich29werenovel.The141miRNAsweremappedontoallchromosomesexcepttheYchromosomeand2/3ofthesemiRNAgenesexistasclusters.sim;#xA0;70%ofthesemiRNAgeneswerelocatedinintronicorintergenicregions,whereastheremainingmiRNAswerederivedfromexonicsequences.WefurthervalidatedtheseclonedmiRNAsbyexaminingtheirexpressioninmultiplemouseorgansincludingdevelopingtestesandalsoinpurifiedspermatogeniccellsusingsemi-quantitativePCRanalyses.Ourexpressionprofilingassaysrevealedthat60%ofthetestis-expressedmiRNAswereubiquitouslyexpressedandtheremainingareeitherpreferentially(35%)orexclusively(5%)expressedinthetestis.WealsoobservedalackofstrandselectionduringtesticularmiRNAbiogenesis,characterizedbypairedexpressionofboththe5prime;strandsand3prime;strandsderivedfromthesameprecursormiRNAs.ThepresentworkidentifiednumerousmiRNAspreferentiallyorexclusivelyexpressedinthetestis,whichwouldbeinterestingtargetsforfurtherfunctionalstudies./p/div