Antisense-Mediated Transcript Knockdown Triggers Premature Transcription Termination.
Mol Cell
; 77(5): 1044-1054.e3, 2020 03 05.
Article
em En
| MEDLINE
| ID: mdl-31924448
Antisense oligonucleotides (ASOs) that trigger RNase-H-mediated cleavage are commonly used to knock down transcripts for experimental or therapeutic purposes. In particular, ASOs are frequently used to functionally interrogate long noncoding RNAs (lncRNAs) and discriminate lncRNA loci that produce functional RNAs from those whose activity is attributable to the act of transcription. Transcription termination is triggered by cleavage of nascent transcripts, generally during polyadenylation, resulting in degradation of the residual RNA polymerase II (Pol II)-associated RNA by XRN2 and dissociation of elongating Pol II. Here, we show that ASOs act upon nascent transcripts and, consequently, induce premature transcription termination downstream of the cleavage site in an XRN2-dependent manner. Targeting the transcript 3' end with ASOs, however, allows transcript knockdown while preserving Pol II association with the gene body. These results demonstrate that the effects of ASOs on transcription must be considered for appropriate experimental and therapeutic use of these reagents.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
RNA Mensageiro
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Precursores de RNA
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Cromatina
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Oligonucleotídeos Antissenso
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Estabilidade de RNA
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Terminação da Transcrição Genética
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article