DNA ultra-sensitive quantification, a technology for studying HIV unintegrated linear DNA.

Autor:	Roux HM; Université Paris Cité, Institut Cochin, INSERM U1016, CNRS, UMR8104, 75014 Paris, France., Figueiredo S; Université Paris Cité, Institut Cochin, INSERM U1016, CNRS, UMR8104, 75014 Paris, France., Sareoua L; Université Paris Cité, Institut Cochin, INSERM U1016, CNRS, UMR8104, 75014 Paris, France., Salmona M; Université Paris Cité, Paris, France.; INSERM U976, Paris, France.; Assistance Publique Hôpitaux de Paris, Hôpital Saint Louis, Laboratoire de Virologie, Paris, France., Hamroune J; Université Paris Cité, Institut Cochin, INSERM U1016, CNRS, UMR8104, 75014 Paris, France., Adoux L; Université Paris Cité, Institut Cochin, INSERM U1016, CNRS, UMR8104, 75014 Paris, France., Migraine J; INSERM U1259, Tours, France., Hance A; INSERM U941, Paris, France., Clavel F; Université Paris Cité, Paris, France.; Assistance Publique Hôpitaux de Paris, Hôpital Saint Louis, Laboratoire de Virologie, Paris, France., Cheynier R; Université Paris Cité, Institut Cochin, INSERM U1016, CNRS, UMR8104, 75014 Paris, France., Dutrieux J; Université Paris Cité, Institut Cochin, INSERM U1016, CNRS, UMR8104, 75014 Paris, France.; Viral DNA Integration and Chromatin Dynamics Network (DyNAVir), France.
Jazyk:	angličtina
Zdroj:	Cell reports methods [Cell Rep Methods] 2023 Apr 05; Vol. 3 (4), pp. 100443. Date of Electronic Publication: 2023 Apr 05 (Print Publication: 2023).
DOI:	10.1016/j.crmeth.2023.100443
Abstrakt:	Unintegrated HIV DNA represents between 20% and 35% of the total viral DNA in infected patients. Only the linear forms (unintegrated linear DNAs [ULDs]) can be substrates for integration and for the completion of a full viral cycle. In quiescent cells, these ULDs may be responsible for pre-integrative latency. However, their detection remains difficult due to the lack of specificity and sensitivity of existing techniques. We developed an ultra-sensitive, specific, and high-throughput technology for ULD quantification called DUSQ (DNA ultra-sensitive quantification) combining linker-mediated PCR and next-generation sequencing (NGS) using molecular barcodes. Studying cells with different activity levels, we determined that the ULD half-life goes up to 11 days in resting CD4 + T cells. Finally, we were able to quantify ULDs in samples from patients infected with HIV-1, providing a proof of concept for the use of DUSQ in vivo to track pre-integrative latency. DUSQ can be adapted to the detection of other rare DNA molecules. Competing Interests: The authors declare no competing interests. (© 2023 The Authors.)
Databáze:	MEDLINE
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