Minimal impact of ZAP on lentiviral vector production and transduction efficiency

Helin Sertkaya, Laura Hidalgo Lumbreras, Mattia Ficarelli, Dorota Kmiec, Adrian Signell, Sadfer Ali, Hannah Parker, Harry Wilson, Stuart Neil, Michael Malim, Conrad A. Vink, Chad Swanson*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)
30 Downloads (Pure)

Abstract

The antiviral protein ZAP binds CpG dinucleotides in viral RNA to inhibit replication. This has likely led to the CpG suppression observed in many RNA viruses, including retroviruses. Sequences added to retroviral vector genomes, such as internal promoters, transgenes or regulatory elements, substantially increase CpG abundance. Because these CpGs could allow retroviral vector RNA to be targeted by ZAP, we analyzed whether it restricts vector production, transduction efficiency and transgene expression. Surprisingly, even though CpG-high HIV-1 was efficiently inhibited by ZAP in HEK293T cells, depleting ZAP did not substantially increase lentiviral vector titre using several packaging and genome plasmids. ZAP overexpression also did not inhibit lentiviral vector titre. In addition, decreasing CpG abundance in a lentiviral vector genome did not increase its titre and a gammaretroviral vector derived from murine leukemia virus was not substantially restricted by ZAP. Overall, we show that the increased CpG abundance in retroviral vectors relative to the wild type retroviruses they are derived from does not intrinsically sensitize them to ZAP. Further understanding of how ZAP specifically targets transcripts to inhibit their expression may allow the development of CpG sequence contexts that efficiently recruit or evade this antiviral system.
Original languageEnglish
Pages (from-to)147-157
Number of pages11
JournalMolecular Therapy - Methods and Clinical Development
Volume23
DOIs
Publication statusPublished - 10 Dec 2021

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