City of Hope

Elizabeth Epps

John J Rossi

John C Burnett

Provided herein are, inter alia, oligonucleotides, kits, and methods useful for increasing lentiviral titers.

Splice inhibiting oligonucleotides

MicroRNAs embedded within an intron, which are called 'mirtrons,' can be used as a platform for expressing one or more shRNA or miRNA mimics in a lentiviral vector. The inventors developed a strategy to improve lentiviral titering by reducing the production of shRNA/miRNA from the vector during packaging through the introduction of splice-inhibiting antisense oligonucleotides during vector packaging, which inhibit the splicing of the mirtron and subsequent processing of the shRNAs/miRNAs. In an aspect is provided a kit comprising an oligonucleotide comprising a mirtron splice site binding sequence and a lentiviral packaging system. In an aspect is provided a method for producing a lentivirus. The method comprises the step of transfecting a cell with an oligonucleotide comprising a mirtron splice site binding sequence and a lentiviral packaging system; thereby producing the lentivirus.

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Mission Viejo, CA, United States of America

Elizabeth Epps