Targeted editing of the PSIP1 gene encoding LEDGF/p75 protects cells against HIV infection

Scientific Reports
Yulia LampiRik Gijsbers

Abstract

To fulfill a productive infection cycle the human immunodeficiency virus (HIV) relies on host-cell factors. Interference with these co-factors holds great promise in protecting cells against HIV infection. LEDGF/p75, encoded by the PSIP1 gene, is used by the integrase (IN) protein in the pre-integration complex of HIV to bind host-cell chromatin facilitating proviral integration. LEDGF/p75 depletion results in defective HIV replication. However, as part of its cellular function LEDGF/p75 tethers cellular proteins to the host-cell genome. We used site-specific editing of the PSIP1 locus using CRISPR/Cas to target the aspartic acid residue in position 366 and mutated it to asparagine (D366N) to disrupt the interaction with HIV IN but retain LEDGF/p75 cellular function. The resulting cell lines demonstrated successful disruption of the LEDGF/p75 HIV-IN interface without affecting interaction with cellular binding partners. In line with LEDGF/p75 depleted cells, D366N cells did not support HIV replication, in part due to decreased integration efficiency. In addition, we confirm the remaining integrated provirus is more silent. Taken together, these results support the potential of site-directed CRISPR/Cas9 mediated knock-in to rend...Continue Reading

References

Sep 29, 1988·Nature·D Baltimore
Aug 31, 2002·Cell·Astrid R W SchröderFrederic Bushman
Oct 31, 2002·The Journal of Biological Chemistry·Peter CherepanovZeger Debyser
May 18, 2004·Current Opinion in Cell Biology·Kristian E Baker, Roy Parker
Oct 12, 2004·The Journal of Biological Chemistry·Manuel LlanoEric M Poeschla
May 17, 2005·Nature Structural & Molecular Biology·Peter CherepanovAlan Engelman
Nov 2, 2005·Proceedings of the National Academy of Sciences of the United States of America·Peter CherepanovAlan Engelman
Nov 29, 2005·Nature Medicine·Angela CiuffiFrederic Bushman
Jun 1, 2006·Journal of Cell Science·Goedele N MaertensAlan Engelman
Sep 9, 2006·Science·Manuel LlanoEric M Poeschla
Dec 2, 2006·Journal of Molecular Biology·Katrien BusschotsFrauke Christ
Aug 3, 2007·Journal of Molecular Biology·Koen BartholomeeusenFrauke Christ
Sep 12, 2007·Annual Review of Medicine·Luc GeeraertRoger J Pomerantz
May 14, 2008·Journal of the American Chemical Society·Mary-Catherine BowmanChristian Melander
Jul 5, 2008·Cancer Cell·Akihiko Yokoyama, Michael L Cleary
Feb 14, 2009·The New England Journal of Medicine·Gero HütterEckhard Thiel
Feb 27, 2009·The Journal of Biological Chemistry·Koen BartholomeeusenJan De Rijck
Sep 29, 2009·Antiviral Research·Catherine S Adamson, Eric O Freed
Oct 29, 2009·Current Topics in Medicinal Chemistry·Samantha BarichievyMarc S Weinberg
Mar 3, 2010·Molecular Therapy : the Journal of the American Society of Gene Therapy·Rik GijsbersZeger Debyser
Aug 7, 2010·Current Opinion in Critical Care·John Lekakis, Ignatios Ikonomidis
Sep 3, 2010·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Marina Núñez
Jan 26, 2011·Microbes and Infection·Chenglai XiaShuwen Liu
Jun 2, 2011·Annals of Internal Medicine·Carl W Dieffenbach, Anthony S Fauci
Feb 16, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Sofie VetsRik Gijsbers
Oct 3, 2012·Cold Spring Harbor Perspectives in Medicine·James A Hoxie, Carl H June
Oct 11, 2012·Retrovirology·Rik SchrijversRik Gijsbers
Dec 12, 2012·Virology·Frauke Christ, Zeger Debyser
Oct 26, 2013·Nature Protocols·F Ann RanFeng Zhang

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Citations

Sep 23, 2020·Journal of Cellular Physiology·Baohong Zhang
Sep 15, 2020·Frontiers in Cellular and Infection Microbiology·Chantelle L AhlenstielAnthony D Kelleher
Apr 9, 2021·Transgenic Research·Abozar GhorbaniG Gregory Neely

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Methods Mentioned

BETA
PCR
transfection
coIP
Co-immunoprecipitation
flow cytometry
FCS
Fluorescence Activated Cell Sorting
Protein Assay
electrophoresis
ELISA

Software Mentioned

MCF
Integrative Genomics Viewer ( IGV )
iQ5 Optical System
Bowtie2
GraphPad Prism
ea
fastQC
fastq
utils

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