METHODS, SYSTEMS AND DEVICES FOR SELECTION AND GENERATION OF GENOME EDITED CLONES

• Main Authors: LAVIEU, Gregory G, MOCCIARO, Annamaria, MCEWEN, Jason, RAMENANI, Ravi K, GUAN RADSTROM, Xiao, DYCK, Patricia A, KURZ, Volker L, SOUMILLON, Magali, NEVILL, J. Tanner
• Format: Patent
• Language: English; French; German
• Published: 23.01.2019
• Subjects: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; PERFORMING OPERATIONS; PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL; PHYSICS; TESTING; TRANSPORTING

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.