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      γ-TRIS: a graph-algorithm for comprehensive identification of vector genomic insertion sites

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          Abstract

          Summary

          Retroviruses and their vector derivatives integrate semi-randomly in the genome of host cells and are inherited by their progeny as stable genetic marks. The retrieval and mapping of the sequences flanking the virus-host DNA junctions allows the identification of insertion sites in gene therapy or virally infected patients, essential for monitoring the evolution of genetically modified cells in vivo. However, since ∼30% of insertions land in low complexity or repetitive regions of the host cell genome, they cannot be correctly assigned and are currently discarded, limiting the accuracy and predictive power of clonal tracking studies. Here, we present γ-TRIS, a new graph-based genome-free alignment tool for identifying insertion sites even if embedded in low complexity regions. By using γ-TRIS to reanalyze clinical studies, we observed improvements in clonal quantification and tracking.

          Availability and implementation

          Source code at https://bitbucket.org/bereste/g-tris.

          Supplementary information

          Supplementary data are available at Bioinformatics online.

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          Most cited references11

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          Search and clustering orders of magnitude faster than BLAST.

          Biological sequence data is accumulating rapidly, motivating the development of improved high-throughput methods for sequence classification. UBLAST and USEARCH are new algorithms enabling sensitive local and global search of large sequence databases at exceptionally high speeds. They are often orders of magnitude faster than BLAST in practical applications, though sensitivity to distant protein relationships is lower. UCLUST is a new clustering method that exploits USEARCH to assign sequences to clusters. UCLUST offers several advantages over the widely used program CD-HIT, including higher speed, lower memory use, improved sensitivity, clustering at lower identities and classification of much larger datasets. Binaries are available at no charge for non-commercial use at http://www.drive5.com/usearch.
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            HIV latency. Specific HIV integration sites are linked to clonal expansion and persistence of infected cells.

            The persistence of HIV-infected cells in individuals on suppressive combination antiretroviral therapy (cART) presents a major barrier for curing HIV infections. HIV integrates its DNA into many sites in the host genome; we identified 2410 integration sites in peripheral blood lymphocytes of five infected individuals on cART. About 40% of the integrations were in clonally expanded cells. Approximately 50% of the infected cells in one patient were from a single clone, and some clones persisted for many years. There were multiple independent integrations in several genes, including MKL2 and BACH2; many of these integrations were in clonally expanded cells. Our findings show that HIV integration sites can play a critical role in expansion and persistence of HIV-infected cells. Copyright © 2014, American Association for the Advancement of Science.
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              Gene therapy returns to centre stage.

              Recent clinical trials of gene therapy have shown remarkable therapeutic benefits and an excellent safety record. They provide evidence for the long-sought promise of gene therapy to deliver 'cures' for some otherwise terminal or severely disabling conditions. Behind these advances lie improved vector designs that enable the safe delivery of therapeutic genes to specific cells. Technologies for editing genes and correcting inherited mutations, the engagement of stem cells to regenerate tissues and the effective exploitation of powerful immune responses to fight cancer are also contributing to the revitalization of gene therapy.
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                Author and article information

                Contributors
                Role: Associate Editor
                Journal
                Bioinformatics
                Bioinformatics
                bioinformatics
                Bioinformatics
                Oxford University Press
                1367-4803
                1367-4811
                March 2020
                07 October 2019
                07 October 2019
                : 36
                : 5
                : 1622-1624
                Affiliations
                [1 ] San Raffaele Telethon Institute for Gene Therapy (SR-Tiget) , IRCCS San Raffaele Scientific Institute, via Olgettina 60, 20132, Milan, Italy
                [2 ] Università degli Studi di Milano Bicocca, Dipartimento di Informatica Sistemistica e Comunicazione (DiSCO) , Viale Sarca, 336, 20126, Milano, Italy
                [3 ] National Research Council, Institute for Biomedical Technologies , Via Fratelli Cervi, 93, 20090, Segrate, Italy
                Author notes

                The authors wish it to be known that, in their opinion, Andrea Calabria, Stefano Beretta and Ivan Merelli should be regarded as Joint First Authors.

                To whom correspondence should be addressed. E-mail: montini.eugenio@ 123456hsr.it
                Author information
                http://orcid.org/0000-0003-3515-3384
                Article
                btz747
                10.1093/bioinformatics/btz747
                7703754
                31589304
                bf842822-17dc-4fc4-a99e-94552a161770
                © The Author(s) 2019. Published by Oxford University Press.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

                History
                : 29 October 2018
                : 18 September 2019
                : 01 October 2019
                Page count
                Pages: 3
                Funding
                Funded by: Telethon Foundation, DOI 10.13039/501100002426;
                Award ID: TGT11D1
                Award ID: TGT16B01
                Award ID: TGT16B03
                Funded by: ISCRA;
                Award ID: HP10CEUWXF
                Funded by: Giovani Ricercatori;
                Award ID: GR-2016-02363681
                Funded by: University and Research flagship initiative Interomics;
                Categories
                Applications Note
                Genome Analysis

                Bioinformatics & Computational biology
                Bioinformatics & Computational biology

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