We apply long-read nanopore sequencing and a new tool, TLDR (Transposons from Long Dirty Reads), to directly infer CpG methylation of new and extant human transposable element (TE) insertions in hippocampus, heart, and liver, as well as paired tumour and nontumour liver. Whole genome TLDR analysis greatly facilitates studies of TE biology as complete insertion sequences and their epigenetic modifications are readily obtainable. Main Transposable elements (TEs) pervade our genomic architecture and broadly influence human biology and disease 1. Recently, Oxford Nanopore Technologies (ONT) long-read DNA sequencing has enabled telomere-to-telomere chromosome assembly at base pair resolution, including of high copy number TEs previously refractory to short-read mapping 2-4. While most evolutionarily older TEs have accumulated sufficient nucleotide diversity to be uniquely identified, recent TE insertions are often indistinguishable from their source elements when assayed with short-read approaches. Each diploid human genome contains 80-100 potentially mobile long interspersed element 1 (LINE-1) copies, referred to here as L1Hs (L1 Human-specific) 5,6. L1Hs elements encode proteins required to retrotranspose 7 in cis, and to trans mobilise Alu and SVA retrotransposons and processed mRNAs 8-10. While the reference genome assembly contains thousands of human-specific TE copies, the vast majority of germline polymorphic 1
1 We apply long-read nanopore sequencing and a new tool, TLDR (Transposons from Long 2 Dirty Reads), to directly infer CpG methylation of new and extant human transposable 3 element (TE) insertions in hippocampus, heart, and liver, as well as paired tumour and non-4 tumour liver. Whole genome TLDR analysis greatly facilitates studies of TE biology as 5 complete insertion sequences and their epigenetic modifications are readily obtainable. 6 7 Main 8Transposable elements (TEs) pervade our genomic architecture and broadly influence 9 human biology and disease 1 . Recently, Oxford Nanopore Technologies (ONT) long-read 10
A recent study proposed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) hijacks the LINE-1 (L1) retrotransposition machinery to integrate into the DNA of infected cells. If confirmed, this finding could have significant clinical implications. Here, we apply deep (>50×) long-read Oxford Nanopore Technologies (ONT) sequencing to HEK293T cells infected with SARS-CoV-2, and do not find the virus integrated into the genome. By examining ONT data from separate HEK293T cultivars, we completely resolve 78 L1 insertions arising
in vitro
in the absence of L1 overexpression systems. ONT sequencing applied to hepatitis B virus (HBV) positive liver cancer tissues located a single HBV insertion. These experiments demonstrate reliable resolution of retrotransposon and exogenous virus insertions via ONT sequencing. That we find no evidence of SARS-CoV-2 integration suggests such events are, at most, extremely rare
in vivo
, and therefore are unlikely to drive oncogenesis or explain post-recovery detection of the virus.
A recent study proposed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) hijacks the LINE-1 (L1) retrotransposition machinery to integrate into the DNA of infected cells. If confirmed, this finding could have significant clinical implications. Here, we applied deep (>50x) long-read Oxford Nanopore Technologies (ONT) sequencing to HEK293T cells infected with SARS-CoV-2, and did not find any evidence of the virus existing as DNA. By examining ONT data from separate HEK293T cultivars, we resolved the complete sequences of 78 L1 insertions arising in vitro in the absence of L1 overexpression systems. ONT sequencing applied to hepatitis B virus (HBV) positive liver cancer tissues located a single HBV insertion. These experiments demonstrate reliable resolution of retrotransposon and exogenous virus insertions via ONT sequencing. That we found no evidence of SARS-CoV-2 integration suggests such events in vivo are highly unlikely to drive later oncogenesis or explain post-recovery detection of the virus.
Pseudomonas baetica strain a390T is the type strain of this recently described species and here we present its high-contiguity draft genome. To celebrate the 16th International Conference on Pseudomonas, the genome of P. baetica strain a390T was sequenced using a unique combination of Ion Torrent semiconductor and Oxford Nanopore methods as part of a collaborative community-led project. The use of high-quality Ion Torrent sequences with long Nanopore reads gave rapid, high-contiguity and -quality, 16-contig genome sequence. Whole genome phylogenetic analysis places P. baetica within the P. koreensis clade of the P. fluorescens group. Comparison of the main genomic features of P. baetica with a variety of other Pseudomonas spp. suggests that it is a highly adaptable organism, typical of the genus. This strain was originally isolated from the liver of a diseased wedge sole fish, and genotypic and phenotypic analyses show that it is tolerant to osmotic stress and to oxytetracycline.
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