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Widespread formation of double-stranded RNA in testis

Lookup NU author(s): Dr Andreas Werner, James Clark, Calum Samaranayake, John CasementORCiD, Dr Hany Zinad, Dr Dr Shaymaa Sadeq, Dr Surar Al-Hashimi



This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0).


Testis express a highly complex transcriptome including RNAs that potentially hybridize to form double-stranded RNA (dsRNA). We isolated dsRNA using the monoclonal J2 antibody and deep- sequenced the enriched samples from testis of juvenile Dicer1 knock-out mice, age-matched controls and adult animals. Comparison of our dataset with recently published data from mouse liver revealed that the dsRNA transcriptome in testis is markedly different from liver: In testis, dsRNA-forming transcripts derive from mRNAs including promoter and immediate downstream region, whereas in somatic cells they originate more often from introns and intergenic transcription. The genes that generate dsRNA are significantly expressed in isolated male germ cells with particular enrichment in pachytene spermatocytes. DsRNA-formation is lower on the sex (X and Y) chromosomes. The dsRNA transcriptome is significantly less complex in juvenile mice as compared to adult controls and, possibly as a consequence, the knock-out of Dicer1 had only a minor effect of the total number of transcript peaks associated with dsRNA. The comparison between dsRNA-associated genes in testis and liver with a reported set of genes that produce endogenous siRNAs revealed a significant overlap in testis but not in liver. Testis dsRNA also significantly associates with natural antisense genes–again, this feature was not observed in liver. These findings point to a testis-specific mechanism involving natural antisense transcripts and the formation of dsRNAs that feed into the RNA interference pathway, possibly to mitigate the mutagenic impacts of recombination and transposon mobilization.

Publication metadata

Author(s): Werner A, Clark JE, Samaranayake C, Casement J, Zinad HS, Sadeq S, Al-Hashimi S, Smith M, Kotaja N, Mattick JS

Publication type: Article

Publication status: Published

Journal: Genome Research

Year: 2021

Volume: 31

Pages: 1174-1186

Print publication date: 01/07/2021

Online publication date: 22/06/2021

Acceptance date: 03/06/2021

Date deposited: 04/06/2021

ISSN (print): 1088-9051

ISSN (electronic): 1549-5469

Publisher: Cold Spring Harbor Laboratory Press


DOI: 10.1101/gr.265603.120


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