eprintid: 10121414 rev_number: 21 eprint_status: archive userid: 608 dir: disk0/10/12/14/14 datestamp: 2021-02-11 09:25:49 lastmod: 2022-09-06 10:34:08 status_changed: 2021-02-11 09:25:49 type: article metadata_visibility: show creators_name: Counsell, JR creators_name: De Brabandere, G creators_name: Karda, R creators_name: Moore, M creators_name: Greco, A creators_name: Bray, A creators_name: Diaz, JA creators_name: Perocheau, DP creators_name: Mock, U creators_name: Waddington, SN title: Re-structuring lentiviral vectors to express genomic RNA via cap-dependent translation ispublished: pub subjects: GOSH divisions: UCL divisions: B02 divisions: C10 divisions: D16 divisions: D11 divisions: G12 divisions: D13 divisions: G23 note: © 2020 The Authors. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). abstract: Lentiviral (LV) vectors based on human immunodeficiency virus type I (HIV-1) package two copies of their single-stranded RNA into vector particles. Normally, this RNA genome is reverse transcribed into a double-stranded DNA provirus that integrates into the cell genome, providing permanent gene transfer and long-term expression. Integration-deficient LV vectors have been developed to reduce the frequency of genomic integration and thereby limit their persistence in dividing cells. Here, we describe optimization of a reverse-transcriptase-deficient LV vector, which enables direct translation of LV RNA genomes upon cell entry, for transient expression of vector payloads as mRNA without a DNA intermediate. We have engineered a novel LV genome arrangement in which HIV-1 sequences are removed from the 5′ end, to enable ribosomal entry from the 5′ 7-methylguanylate cap for efficient translation of the vector payload. We have shown that this LV-mediated mRNA delivery platform provides transient transgene expression in vitro and in vivo. This has a potential application in gene and cell therapy scenarios requiring temporary payload expression in cells and tissues that can be targeted with pseudotyped LV vectors. date: 2021-03-12 date_type: published official_url: https://doi.org/10.1016/j.omtm.2020.12.005 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1845466 doi: 10.1016/j.omtm.2020.12.005 lyricists_name: Antinao Diaz, Juan lyricists_name: Counsell, John lyricists_name: Greco, Antonio lyricists_name: Karda, Rajvinder lyricists_name: Perocheau, Dany lyricists_name: Waddington, Simon lyricists_id: JFANT33 lyricists_id: JRCOU46 lyricists_id: AGREC71 lyricists_id: RKARD12 lyricists_id: DPERO72 lyricists_id: WADDI73 actors_name: Kalinowski, Damian actors_id: DKALI47 actors_role: owner full_text_status: public publication: Molecular Therapy - Methods and Clinical Development volume: 20 pagerange: 357-365 citation: Counsell, JR; De Brabandere, G; Karda, R; Moore, M; Greco, A; Bray, A; Diaz, JA; ... Waddington, SN; + view all <#> Counsell, JR; De Brabandere, G; Karda, R; Moore, M; Greco, A; Bray, A; Diaz, JA; Perocheau, DP; Mock, U; Waddington, SN; - view fewer <#> (2021) Re-structuring lentiviral vectors to express genomic RNA via cap-dependent translation. Molecular Therapy - Methods and Clinical Development , 20 pp. 357-365. 10.1016/j.omtm.2020.12.005 <https://doi.org/10.1016/j.omtm.2020.12.005>. Green open access document_url: https://discovery-pp.ucl.ac.uk/id/eprint/10121414/1/Karda_Re-structuring%20lentiviral%20vectors%20to%20express%20genomic%20RNA%20via%20cap-dependent%20translation_VoR.pdf