Identification of host essential factors for recombinant AAV transduction of the polarized human airway epithelium.
In: Journal of Virology, Jg. 97 (2023-12-01), Heft 12, S. 1-24
academicJournal
Zugriff:
Recombinant (r)AAV2.5T was selected from the directed evolution of an adeno-associated virus (AAV) capsid library in the human airway epithelium (HAE). The capsid gene of rAAV2.5T is a chimera of the N-terminal unique coding sequence of AAV2 VP1 unique (VP1u) and the VP2- and VP3-coding sequences of AAV5 with a single amino acid mutation of A581T. We conducted two rounds of genome-wide CRISPR guide (g)RNA library screening for host factors limiting rAAV2.5T transduction in HeLa S3 cells. The screening identified several genes that are critical for rAAV2.5T, including the previously reported genes KIAA0319L, TM9SF2, VPS51, and VPS54, as well as a novel gene WDR63. We verified the role of KIAA0319L and WDR63 in rAAV2.5T transduction of polarized HAE by utilizing CRISPR gene knockouts. Although KIAA0319L, a proteinaceous receptor for multiple AAV serotypes, played an essential role in rAAV2.5T transduction of polarized HAE either from the apical or basolateral side, our findings demonstrated that the internalization of rAAV2.5T was independent of KIAA0319L. Importantly, we confirmed that WDR63 is an important player in rAAV2.5T transduction of HAE while not being involved in vector internalization and nuclear entry. Furthermore, we ident ified that the basal stem cells of HAE can be significantly transduced by rAAV2.5T. [ABSTRACT FROM AUTHOR]
Titel: |
Identification of host essential factors for recombinant AAV transduction of the polarized human airway epithelium.
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Autor/in / Beteiligte Person: | Hao, Siyuan ; Zhang, Xiujuan ; Ning, Kang ; Feng, Zehua ; Soo Yeun Park ; Kuz, Cagla Aksu ; McFarlin, Shane ; Richart, Donovan ; Cheng, Fang ; Yan Zhang, Elizabeth ; Zhang-Chen, Aaron ; Yan, Ziying ; Qiu, Jianming |
Zeitschrift: | Journal of Virology, Jg. 97 (2023-12-01), Heft 12, S. 1-24 |
Veröffentlichung: | 2023 |
Medientyp: | academicJournal |
ISSN: | 0022-538X (print) |
DOI: | 10.1128/jvi.01330-23 |
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