Compositions, methods, modules and instruments for automated nucleic acid-guided nuclease editing in mammalian cells via viral delivery
US11845932B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 31, 2022 |
| Grant date | Dec 19, 2023 |
| Priority date | — |
| Expiry date | May 31, 2042 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2800/80
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Nucleic acid-guided nuclease editing in mammalian cells may include passaging mammalian cells, in an automated closed cell editing instrument, into smaller aggregates when the aggregates exceed 50-300 microns in size. A library of viral particles may be delivered to the mammalian cells at a multiplicity of infection such that each mammalian cell receives one or no viral particle. The library may include viral vectors with an editing cassette including a pair of gRNA coding sequence and donor DNA. Conditions may be provided to allow a viral vector of the viral vectors to integrate into the mammalian cells. Enriching for mammalian cells may be done with an integrated viral vector. A nucleic acid-guided nuclease or nuclease fusion or a coding sequence for a nucleic acid-guided nuclease or nuclease fusion may be delivered to the enriched mammalian cells and conditions may be provided to allow editing in the mammalian cells.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.