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Subtractive hybridization identifies novel differentially expressed ncRNA species in EBV-infected human B cells.

Abstract：

:Non-protein-coding RNAs (ncRNAs) fulfill a wide range of cellular functions from protein synthesis to regulation of gene expression. Identification of novel regulatory ncRNAs by experimental approaches commonly includes the generation of specialized cDNA libraries encoding small ncRNA species. However, such identification is severely hampered by the presence of constitutively expressed and highly abundant 'house-keeping' ncRNAs, such as ribosomal RNAs, small nuclear RNAs or transfer RNAs. We have developed a novel experimental strategy, designated as subtractive hybridization of ncRNA transcripts (SHORT) to specifically select and amplify novel regulatory ncRNAs, which are only expressed at certain stages or under specific growth conditions of cells. The method is based on the selective subtractive hybridization technique, formerly applied to the detection of differentially expressed mRNAs. As a model system, we applied SHORT to Epstein-Barr virus (EBV) infected human B cells. Thereby, we identified 21 novel as well as previously reported ncRNA species to be up-regulated during virus infection. Our method will serve as a powerful tool to identify novel functional ncRNAs acting as genetic switches in the regulation of fundamental cellular processes such as development, tissue differentiation or disease.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Mrázek J,Kreutmayer SB,Grässer FA,Polacek N,Hüttenhofer A

doi

10.1093/nar/gkm244

subject

Has Abstract

pub_date

2007-01-01 00:00:00

pages

e73

issue

10

eissn

0305-1048

issn

1362-4962

pii

gkm244

journal_volume

35

pub_type

杂志文章

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