Article Dans Une Revue Biochimie Année : 2024

Evaluation of microRNA variant maturation prior to genome edition

Evaluation de la maturation de variants de microARN préalablement à l'édition génomique

Résumé

Assessment of the functionality of individual microRNA/target sites is a crucial issue. Genome editing techniques should theoretically permit a fine functional exploration of such interactions, allowing the mutation of microRNAs or individual binding sites in a complete in vivo setting, therefore abrogating or restoring individual interactions on demand. A major limitation to this experimental strategy is the influence of microRNA sequence on its accumulation level, which introduces a confounding effect when assessing phenotypic rescue by compensatorily mutated microRNA and target site. Here we describe a simple assay to identify microRNA variants most likely to accumulate at wild-type levels even though their sequence has been mutated. In this assay, quantification of a reporter construct in cultured cells predicts the efficiency of an early biogenesis step, the Drosha-dependent cleavage of microRNA precursors, which appears to be a major determinant of microRNA accumulation in our variant collection. This system allowed the generation of a mutant Drosophila strain expressing a bantam microRNA variant at wild-type levels.

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hal-04661180 , version 1 (24-07-2024)

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Isabelle Busseau, Sophie Mockly, Élisabeth Houbron, Hedi Somaï, Hervé Seitz. Evaluation of microRNA variant maturation prior to genome edition. Biochimie, 2024, 217, pp.86-94. ⟨10.1016/j.biochi.2023.06.007⟩. ⟨hal-04661180⟩
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