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.

中文翻译：

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反义介导的转录敲低触发转录终止。

触发RNase-H介导的切割的反义寡核苷酸（ASO）通常用于敲除转录本，以用于实验或治疗目的。特别是，ASO通常用于功能性询问长的非编码RNA（lncRNA），并从可能归因于其转录活性的分子中区分出产生功能性RNA的lncRNA基因座。通常在聚腺苷酸化过程中，通过新生转录物的切割触发转录终止，从而导致XRN2降解残留的RNA聚合酶II（Pol II）相关的RNA，并破坏延伸的Pol II。在这里，我们表明ASOs对新生的转录物起作用，并因此以XRN2依赖性的方式诱导切割位点下游的过早转录终止。但是，将转录本3'末端定位为ASO 在保留Pol II与基因体的关联的同时，允许转录物敲低。这些结果表明，对于这些试剂的适当实验和治疗用途，必须考虑ASO对转录的影响。