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Quantitative assessment reveals the dominance of duplicated sequences in germline-derived extrachromosomal circular DNA
Journal article   Open access   Peer reviewed

Quantitative assessment reveals the dominance of duplicated sequences in germline-derived extrachromosomal circular DNA

Lila Mouakkad-Montoya, Michael M Murata, Arvis Sulovari, Ryusuke Suzuki, Beth Osia, Anna Malkova, Makoto Katsumata, Armando E Giuliano, Evan E Eichler and Hisashi Tanaka
Proceedings of the National Academy of Sciences - PNAS, Vol.118(47), p.1
11/23/2021
DOI: 10.1073/pnas.2102842118
PMCID: PMC8617514
PMID: 34789574
url
https://doi.org/10.1073/pnas.2102842118View
Published (Version of record) Open Access

Abstract

Extrachromosomal circular DNA (eccDNA) originates from linear chromosomal DNA in various human tissues under physiological and disease conditions. The genomic origins of eccDNA have largely been investigated using in vitro-amplified DNA. However, in vitro amplification obscures quantitative information by skewing the total population stoichiometry. In addition, the analyses have focused on eccDNA stemming from single-copy genomic regions, leaving eccDNA from multicopy regions unexamined. To address these issues, we isolated eccDNA without in vitro amplification (naïve small circular DNA, nscDNA) and assessed the populations quantitatively by integrated genomic, molecular, and cytogenetic approaches. nscDNA of up to tens of kilobases were successfully enriched by our approach and were predominantly derived from multicopy genomic regions including segmental duplications (SDs). SDs, which account for 5% of the human genome and are hotspots for copy number variations, were significantly overrepresented in sperm nscDNA, with three times more sequencing reads derived from SDs than from the entire single-copy regions. SDs were also overrepresented in mouse sperm nscDNA, which we estimated to comprise 0.2% of nuclear DNA. Considering that eccDNA can be integrated into chromosomes, germline-derived nscDNA may be a mediator of genome diversity.
Animals Chromosomes DNA DNA Copy Number Variations DNA, Circular Genome, Human Germ Cells HeLa Cells Humans Male Mice Mice, Inbred C57BL Segmental Duplications, Genomic Spermatozoa

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