Methods, systems and devices for selection and generation of genome edited clones

Methods are described herein for isolating clonal populations of cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual cells (or precursors...

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Bibliographische Detailangaben
Hauptverfasser:	Soumillon, Magali, Lavieu, Gregory G, Kurz, Volker L. S, Dyck, Patricia A, Guan Radstrom, Xiao, Ramenani, Ravi K, McEwen, Jason M, Nevill, J. Tanner, Mocciaro, Annamaria
Format:	Patent
Sprache:	eng
Schlagworte:	APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY BEER BIOCHEMISTRY CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE CHEMISTRY COMPOSITIONS THEREOF CULTURE MEDIA ENZYMOLOGY INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLIDMATERIALS OR FLUIDS MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROMSOLID MATERIALS OR FLUIDS MEASURING METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PERFORMING OPERATIONS PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL PHYSICS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATICTABLES OR JIGS SPIRITS TESTING TRANSPORTING VINEGAR WINE
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Beschreibung
Zusammenfassung:	Methods are described herein for isolating clonal populations of cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the individual cells into respective clonal populations of cells; and detecting, in one or more cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of cells. Also described are methods of performing genome editing within a microfluidic device, and compositions comprising one or more clonal populations of cells generated according to the methods disclosed herein.