Locked-nucleotide analog antagonists (LNAA) to four varicella zoster virus small noncoding RNA (VZVsncRNA 10–13) derived from the mRNA of the open reading frame (ORF) 61 gene individually reduce VZV replication in epithelial cells and fibroblasts. To study the potential roles VZVsncRNA 10–13 have in neuronal infection we generated two recombinant VZV; one in which 8 nucleotides were changed in VZVsncRNA10 without altering the encoded residues of ORF61 (VZVsnc10MUT) and a second containing a 12-nucleotide deletion of the sequence common to VZVsncRNA12 and 13, located in the ORF61 mRNA leader sequence (VZVsnc12-13DEL). Both were developed from a VZV BAC with a green fluorescent protein (GFP) reporter fused to the N terminal of the capsid protein encoded by ORF23. The growth of both mutant VZV in epithelial cells and fibroblasts was similar to that of the parental recombinant virus. Both mutants established productive infections and experimental latency in neurons derived from human embryonic stem cells (hESC). However, neurons that were latently infected with both VZV mutant viruses showed impaired ability to reactivate when given stimuli that successfully reactivated the parental virus. These results suggest that these VZVsncRNA may have a role in VZV latency maintenance and/or reactivation. The extension of these studies and confirmation of such roles could potentially inform the development of a non-reactivating, live VZV vaccine.
|State||Published - 10 May 2022|
Bibliographical noteFunding Information:
Acknowledgments: P.R.K. wishes to acknowledge NEI CORE grant P30 EY08098 and unrestricted support from the Eye & Ear Foundation of Pittsburgh and the Research to Prevent Blindness Inc, NY. P.R.K. and R.S.G. wish to dedicate this manuscript to the memory of our friend and central figure in the study of the molecular virology of VZV, Randall Cohrs. We both owe Randy a great deal for his insightful discussions and important studies that greatly advanced our field.
Funding: This work was supported by an Israel-US Binational Science Foundation award (#2017259) and awards from the National Institutes of Health from NIAID (AI122640 & AI156527) to P.R.K and R.S.G); and an Israel Science Foundation award (#613/20) to R.S.G.
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.
- Human neuron culture
- Non-coding RNA
- Varicella zoster virus