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Template switching mechanism drives the tandem amplification of chromosome 20q11.21 in human pluripotent stem cells

Jason A Halliwell; Duncan Baker; Kim Judge; Michael A Quail; Karen Oliver; Emma Betteridge; Jason Skelton; Peter W Andrews; Ivana Barbaric

2020-07-12 genomics
10.1101/2020.07.11.198382 bioRxiv
Show abstract

Copy number variants (CNVs) are genomic rearrangements implicated in numerous congenital and acquired diseases, including cancer. In human pluripotent stem cells (PSC), the appearance of culture-acquired CNVs prompted concerns for their use in regenerative medicine applications. A particularly common problem in PSC is the occurrence of CNVs in the q11.21 region of chromosome 20. However, the exact mechanisms of origin of this amplicon remains elusive due to the difficulty in delineating its sequence and breakpoints. Here, we used long-range Nanopore sequencing on two examples of this CNV, present as a duplication in one and a triplication in another line. The CNVs were arranged in a head-to-tail orientation in both lines, with sequences of microhomologies flanking or overlapping both the proximal and distal breakpoints. These breakpoint signatures point to a specific mechanism of template switching in CNV formation, with surrounding Alu sequences likely contributing to the instability of this genomic region.

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