Lentiviral vectors for high-titer transduction of primary human cells
AssigneeUniversity of Massachusetts
InventorsJeremy Luban, Kyusik Kim, Sean Matthew McCauley
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Abstract
Aspects of the disclosure relate to packagable RNA constructs with minimal intervening viral sequences. These constructs can be used to generate lentiviral viruses encoding large genes capable of transducing primary human cells.
Claims71
A construct comprising a nucleic acid encoding a promoter operably linked to transgene encoding a packagable vector RNA, the packagable vector RNA comprising 5′- and 3′-terminal repeats (TRs) that flank:
- i. a nucleocapsid protein packaging target site,
- ii. a heterologous nucleic acid insert, and
- iii. minimal intervening viral sequences.
A nucleic acid comprising a heterologous nucleic acid insert flanked by terminal repeats (TRs), wherein between a first terminal repeat and the heterologous nucleic acid sequence are present packaging and nuclear export sequences and minimal intervening viral sequences.
A transcribed nucleic acid encoding a heterologous nucleic acid insert flanked by TRs, wherein between the first TR- and the heterologous nucleic acid sequence, there are sequences that aid in the packaging and nuclear export of the transcribed nucleic acid and minimal intervening viral sequences.
A transcribed nucleic acid encoding a heterologous nucleic acid insert flanked by terminal repeats (TR), wherein between the first terminal repeat region and the heterologous nucleic acid sequence, there are sequences that aid in the packaging and nuclear export of the transcribed nucleic acid and minimal intervening viral sequences.
A construct comprising a promoter operably linked to a nucleic acid encoding a packagable vector RNA, the packagable vector RNA comprising 5′- and 3′-terminal repeats (TRs) that flank:
- i. a nucleocapsid protein packaging target site,
- ii. a heterologous nucleic acid insert that encodes a microRNA, and
- iii. minimal intervening viral sequences.
The construct of any of claims 60-67, wherein the minimal intervening viral sequences are in total up to 350 base pairs in length.
A construct comprising a sequence as set forth in SEQ ID NO:
- 10.
A construct comprising a sequence as set forth in SEQ ID NO:
- 11.
Dependencies are inferred from the claim text.