University of California

Whitehead Institute for Biomedical Research

Berkeley Lights, Inc.

Alexander Marson

Jennifer A Doudna

Theodore Lee Roth

Cristina Puig Saus

Eric Shifrut

Richard A Young

Rudolf Jaenisch

Jeffrey A Bluestone

Antoni Ribas

Ruth Foreman

Brett Chevalier

Gregory G Lavieu

Annamaria Mocciaro

Magali Soumillon

Francis C Szoka

Hayley M Bennett

Steven Lin

Daniel Goodman

David-Huy Nhu Nguyen

Kathrin Schumann

Provided herein are methods and compositions for editing the genome of a human T cell. In some embodiments, a heterologous T cell receptor (TCR)-β chain and a heterologous TCR-α chain are inserted into exon 1 of a TCR subunit constant gene in the genome of the T cell.

Targeted replacement of endogenous T cell receptors

The disclosure provides compositions and methods for modifying a target nucleic acid. In some embodiments, a composition can include a targetable nuclease, a DNA-binding protein, and a donor template comprising a homology directed repair (HDR) template and one or more DNA-binding protein target sequences. In some embodiments, a composition can include a Cas protein, one or more single guide RNAs (sgRNAs), and an anionic polymer.

Compositions and methods for modifying a target nucleic acid

Provided herein are methods and compositions for editing the genome of a cell. In some embodiments, a nucleotide sequence of at least 200 nucleotides in length is inserted into a target region in the genome of a cell.

Targeted non-viral DNA insertions

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.

Methods for selection and generation of genome edited T cells

Provided herein are methods and compositions for editing the genome of a human T cell. In some embodiments, a heterologous T cell receptor (TCR)-&#x3b2; chain and a heterologous TCR-&#x3b1; chain are inserted into exon 1 of a TCR subunit constant gene in the genome of the T cell.

The invention provides compositions and methods of use in reprogramming somatic cells. Compositions and methods of the invention are of use, e.g., for generating or modulating (e.g., enhancing) generation of induced pluripotent stem cells by reprogramming somatic cells. The reprogrammed somatic cells are useful for a number of purposes, including treating or preventing a medical condition in an individual. The invention further provides methods for identifying an agent that reprograms somatic cells to a pluripotent state and/or enhances the speed and/or efficiency of reprogramming. Certain of the compositions and methods relate to modulating the Wnt pathway.

Wnt pathway stimulation in reprogramming somatic cells with nuclear reprogramming factors

WNT pathway stimulation in reprogramming somatic cells with nuclear reprogramming factors

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San Francisco, CA, United States of America

Alexander Marson