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HIV-1 sequences in lentiviral vector genomes can be substantially reduced without compromising transduction efficiency

Sci Rep. 2021-06; 
Helin Sertkaya, Mattia Ficarelli, Nathan P Sweeney, Hannah Parker, Conrad A Vink, Chad M Swanson
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GenParts™ DNA Fragments For pLV-3’RRE, the RRE and SV40 poly(A) sequences were synthesized using GenScript fragment synthesis and inserted into the KpnI and AvrII sites in pLV∆env. pCMV∆R8.91 Get A Quote

摘要

Many lentiviral vectors used for gene therapy are derived from HIV-1. An optimal vector genome would include only the viral sequences required for transduction efficiency and gene expression to minimize the amount of foreign sequence inserted into a patient's genome. However, it remains unclear whether all of the HIV-1 sequence in vector genomes is essential. To determine which viral sequences are required, we performed a systematic deletion analysis, which showed that most of the gag region and over 50% of the env region could be deleted. Because the splicing profile for lentiviral vectors is poorly characterized, we used long-read sequencing to determine canonical and cryptic splice site usage. Deleting speci... More

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