Barcoded Cells Engineered With Heterozygous Genetic Diversity
OwnerRobert W. Sobol, Jay Georgeinventors
Life of this patent
Estimating the term…
Abstract
The present invention provides a barcoded exon tagging and gene disruption platform to create barcoded control, heterozygous gene knockout (KO) and homozygous gene KO panels of diploid human cells for high-throughput, multiplexed genotoxin screens.
Claims12
A method for generating a population of cells, comprising:
- a) providing a plurality of cells;
- b) modifying a first cell by incorporating a first unique barcode cassette having i) a primer sequence, ii) a first unique barcode, and iii) a selectable marker, into the cell's genome in a safe landing zone to form a control cell;
- c) modifying one or more second cells by incorporating a second unique barcode cassette having i) a primer sequence, ii) a second unique barcode different than the first unique barcode, and iii) a selectable marker, in a target gene in the second cell's genome such that the target gene is rendered inactive by the second unique barcode cassette;
- wherein step c) results in the formation of both homozygous and heterozygous cells with respect to the target gene, and wherein steps b) and c) are performed in any order or simultaneously.
A genetically modified cell whose genome has been modified to incorporate a barcode cassette having i) a primer sequence, ii) a barcode, and iii) a selectable marker, such that a target gene is rendered inactive by the barcode cassette, and wherein the cell is heterozygous with respect to the target gene.
A genetically modified cell whose genome has been modified to incorporate a unique barcode cassette having i) a primer sequence, ii) a first unique barcode, and iii) a selectable marker, into the cell's genome in a safe landing zone.
Dependencies are inferred from the claim text.