In Vitro Transcription of Plant miRNA for Structural and Processing Analysis.

Corina Diaz-Quezada, Noe Baruch-Torres, Carlos H Trasviña-Arenas, Luis G Brieba

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Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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4 authors.

Corina Diaz-QuezadaUnidad de Genomica Avanzada (UGA), Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (Cinvestav), Carretera Irapuato-León, Irapuato, Guanajuato, Mexico.

Noe Baruch-TorresDepartment of Biochemistry and Molecular Biology, University of Texas Medical Branch, USA Sealy Center for Structural Biology, University of Texas Medical Branch, Galveston, TX, USA.

Carlos H Trasviña-ArenasCentro de Investigación sobre Envejecimiento, Cinvestav, Unidad Sede Sur, Mexico City, Mexico.

Luis G BriebaUnidad de Genomica Avanzada (UGA), Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (Cinvestav), Carretera Irapuato-León, Irapuato, Guanajuato, Mexico. luis.brieba@cinvestav.mx.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Plant miRNAs are short gene repressors, ranging in length from 20 to 24 nucleotides, that specifically target mRNAs to effectively promote cleavage or impinge translation. Long primary miRNAs (pri-miRNA) from plants fold into double-stranded RNA structures with stem loop structures. These structures are then processed by an RNase type III enzyme and several other proteins. A functional mature miRNA selects its mRNA target through complementary base pairing. Importantly, plant pri-miRNA are variable in size and structure. Here, we describe a protocol to identify plant pri-miRNA processing intermediates by in vitro T7 RNA polymerase transcription and its purification for enzymatic and chemical footprinting as a tool to analyze the peculiarities of plant miRNA processing.

Indexed as

MicroRNAsRNA, PlantRNA Processing, Post-TranscriptionalTranscription, GeneticDNA-Directed RNA PolymerasesNucleic Acid ConformationViral Proteinsbacteriophage T7 RNA polymeraseDNA-Directed RNA PolymerasesMicroRNAsRNA, PlantViral ProteinsDenaturing gel electrophoresisFootprintingmiRNApri-miRNAProcessingSecondary structureT7 RNA polymerase

Identifiers

PMID40380056

DOI10.1093/nar/gky796

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.