Precise excision of HTLV-1 provirus with a designer-recombinase

Rojo-Romanos T, Karpinski J, Millen S, Beschorner N, Simon F, Paszkowski-Rogacz M, Lansing F, Schneider PM, Sonntag J, Hauber J, Thoma-Kreß A, Buchholz F (2023)

Publication Type: Journal article

Publication year: 2023

Journal

Molecular Therapy Cell Press

DOI: 10.1016/j.ymthe.2023.03.014

Abstract

The human T cell leukemia virus type 1 (HTLV-1) is a pathogenic retrovirus that persists as a provirus in the genome of infected cells and can lead to adult T cell leukemia (ATL). Worldwide, more than 10 million people are infected and approximately 5% of these individuals will develop ATL, a highly aggressive cancer that is currently incurable. In the last years, genome editing tools have emerged as promising antiviral agents. In this proof-of-concept study, we use substrate-linked directed evolution (SLiDE) to engineer Cre-derived site-specific recombinases to excise the HTLV-1 proviral genome from infected cells. We identified a conserved loxP-like sequence (loxHTLV) present in the long terminal repeats of the majority of virus isolates. After 181 cycles of SLiDE, we isolated a designer-recombinase (designated RecHTLV), which efficiently recombines the loxHTLV sequence in bacteria and human cells with high specificity. Expression of RecHTLV in human Jurkat T cells resulted in antiviral activity when challenged with an HTLV-1 infection. Moreover, expression of RecHTLV in chronically infected SP cells led to the excision of HTLV-1 proviral DNA. Our data suggest that recombinase-mediated excision of the HTLV-1 provirus represents a promising approach to reduce proviral load in HTLV-1-infected individuals, potentially preventing the development of HTLV-1-associated diseases.

Authors with CRIS profile

Sebastian Millen Institute of Clinical and Molecular Virology Florian Simon Institute of Clinical and Molecular Virology Andrea Thoma-Kreß Institute of Clinical and Molecular Virology

Additional Organisation(s)

FAU Profilzentrum Licht.Materie.Quantentechnologien (FAU LMQ)

Involved external institutions

Universitätsklinikum Carl Gustav Carus Dresden DE Germany (DE) PROVIREX Genome Editing Therapies GmbH DE Germany (DE)

How to cite

APA:

Rojo-Romanos, T., Karpinski, J., Millen, S., Beschorner, N., Simon, F., Paszkowski-Rogacz, M.,... Buchholz, F. (2023). Precise excision of HTLV-1 provirus with a designer-recombinase. Molecular Therapy. https://dx.doi.org/10.1016/j.ymthe.2023.03.014

MLA:

Rojo-Romanos, Teresa, et al. "Precise excision of HTLV-1 provirus with a designer-recombinase." Molecular Therapy (2023).

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