Analysis of Human Small Nucleolar RNAs (snoRNA) and the Development of snoRNA Modulator of Gene Expression Vectors

Motoharu Ono, Kayo Yamada, Fabio Avolio, Michelle S. Scott, Silvana van Koningsbruggen, Geoffrey J. Barton, Angus I. Lamond

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    29 Citations (Scopus)

    Abstract

    Human small nucleolar RNAs (snoRNAs) that copurify with nucleoli isolated from HeLa cells have been characterized. Novel fibrillarin-associated snoRNAs were detected that allowed the creation of a new vector system for the targeted knockdown of one or more genes in mammalian cells. The snoMEN (snoRNA modulator of gene expressioN) vector technology is based on snoRNA HBII-180C, which contains an internal sequence that can be manipulated to make it complementary to RNA targets. Gene-specific knockdowns are demonstrated for endogenous cellular proteins and for G/YFP-fusion proteins. Multiplex snoMEN vectors coexpress multiple snoRNAs in one transcript, targeted either to different genes or to different sites in the same gene. Protein replacement snoMEN vectors can express a single transcript combining cDNA for a tagged protein with introns containing cognate snoRNAs targeted to knockdown the endogenous cellular protein. We foresee applications for snoMEN vectors in basic gene expression research, target validation, and gene therapy.

    Original languageEnglish
    Pages (from-to)1569-1584
    Number of pages16
    JournalMolecular Biology of the Cell
    Volume21
    Issue number9
    DOIs
    Publication statusPublished - 1 May 2010

    Keywords

    • BIOGENESIS
    • CELLS
    • PROTEINS
    • DYNAMICS
    • REVEALS
    • NUCLEAR
    • GENOMES
    • MOUSE

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