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US 2019/0194694 A1Published application
Improved eukaryotic cells for protein manufacturing and methods of making them
AssigneeSelexis S.A.
InventorsNicolas Mermod, Pierre-Olivier Duroy, Sandra Bosshard, Philippe Le Mercier
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Abstract
Disclosed are mammalian cells and mammalian cell lines that have a reduced load of remnants of past viral/retroviral infections and methods of producing and using the same.
Claims34
An engineered cell, preferably of a mammalian cell line such as an engineered CHO cell, including an engineered CHO-K1, comprising:
- a genome of the cell, wherein the genome comprises one or more alterations comprising:
- deletions,
- additions, and/or
- substitutions,
- of one or more nucleic acids in one or more, generally, more than 10, 20, 30, 40, 50, 60, 80, 90 or 100 endogenous retrovirus (ERV) elements which are part of said genome.
2 more elements
A method for improved genome editing comprising:
- (i) introducing a heterologous system for introducing single or double stranded brakes into a target nucleic acid sequence, such as a consensus sequence of an ERV element, preferably an ERV element that is or is from a gag gene into a cell, preferably a cell of interest, and further
- (ii) introducing into the cell,
- (a) heterologous nucleic acid sequences encoding or activating one or more proteins of one or more of the recombination pathways, and/or
- (b) heterologous sequences encoding one or more sequences, such as siRNAs, or proteins suppressing expression of the one or more proteins of one or more of the recombination pathways, wherein said heterologous system of (i) and/or sequences of (ii) (a) and/or (ii) (b) are preferably at least transiently expressed in the cell of interest.
1 more element
A kit comprising:
- (i) in one container at least one non-integrating vector encoding at least a nuclease, such as CRISPR,
- (ii) in the same or a further container one or more guide RNAs or sequences encoding the one or more guide RNAs targeting a motif in a ERV element
- (iii) in same or a further container one or more siRNAs or sequences encoding one or more siRNAs, and
- in a further container instruction of how to provide (i), (ii) and (iii) within a cell.
1 more element
Dependencies are inferred from the claim text.