Methods for selection and generation of genome edited T cells

US11639495B2 · kind B2 · utility

Assignees

• The Regents of the University of California · US
• Berkeley Lights, Inc. · US

Inventors

• Alexander Marson · San Francisco, US
• Gregory G. Lavieu · Vitry-sur-Seine, FR
• Annamaria Mocciaro · San Francisco, US
• Theodore Lee Roth · San Francisco, US
• Magali Soumillon · Berkeley, US
• Hayley M. Bennett · Emeryville, US

Key dates

• Filing date: Jun 27, 2019
• Grant date: May 2, 2023
• Expiry date: Jan 15, 2042

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N27/447
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Methods are described herein for isolating clonal populations of T 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 T cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the T cells into respective clonal populations of T cells; detecting, in one or more T cells of each clonal population, the absence of a cell surface marker that was present in the individual T cells (or precursors thereof); and detecting, in one or more T 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 T cells. Also described are compositions comprising one or more clonal populations of T cells isolated according to the methods disclosed herein.