Inscripta, Inc.

University of Colorado

Andrew Garst

Ryan T Gill

Phillip Belgrader

Don Masquelier

Richard Fox

Aamir Mir

Kyle Seamon

Benjamin Mijts

Tanya Elizabeth Warnecke Lipscomb

Juhan Kim

Bruce Chabansky

Jorge Bernate

Stephen Federowicz

Ramsey Ibrahim Zeitoun

Matthew David Estes

Michael Graige

Eileen Spindler

Marcelo Colika Bassalo

Amy Hiddessen

Charles D Johnson

Tian Tian

Miles Gander

Daniel Held

Robert T Batey

Emily Feldman

Erik Zimmerman

Roman S Iwasaki Cordero

Provided herein are single guide RNAs (sgRNAs) that comprise aptamer sequences and related compositions and methods. Also provided herein are methods of selecting inducible sgRNAs that comprise aptamer sequences.

Compositions and methods of use for small-molecule regulation of CRISPR-Cas9 activity using RNA aptamers

Disclosed herein are nucleic acid-guided nucleases, guide nucleic acids, and targetable nuclease systems, and methods of use. Disclosed herein are engineered non-naturally occurring nucleic acid-guided nucleases, guide nucleic acids, and targetable nuclease systems, and methods of use. Targetable nuclease systems can be used to edit genetic targets, including recursive genetic engineering and trackable genetic engineering methods.

Nucleic acid-guided nucleases

Described herein are method for generating a vector for editing a cell. The method comprises ligating into a vector that encodes a portion of a gRNA a cassette comprising at least one editing cassette, a promoter, and a gene encoding another portion of the gRNA. Upon ligation, the portion of the gRNA from the editing cassette and the other portion of the gRNA are ligated and form a functional gRNA.

CRISPR enabled multiplexed genome engineering

The present disclosure provides instruments, modules and methods for improved detection of edited cells following nucleic acid-guided nuclease genome editing. The disclosure provides improved automated instruments that perform methods&#x2014;including high throughput methods&#x2014;for screening cells that have been subjected to editing and identifying cells that have been properly edited.

Instruments, modules, and methods for improved detection of edited sequences in live cells

Described herein are methods and vectors for rational, multiplexed manipulation of chromosomes within open reading frames (e.g., in protein libraries) or any segment of a chromosome in a cell or population of cells, in which various CRISPR systems are used.

The present disclosure provides automated modules and instruments for improved detection of nuclease genome editing of live cells. The disclosure provides improved modules&#x2014;including high throughput modules&#x2014;for screening cells that have been subjected to editing and identifying and selecting cells that have been properly edited.

Detection of nuclease edited sequences in automated modules and instruments

Described herein are molecules for editing a cell. The molecules described herein generally comprise the following covalently-linked components and a nucleic acid encoding a guide RNA (gRNA) sequence targeting a target region in a cell and a region homologous to the target region comprising a change in sequence relative to the target region.

Provided herein are methods and composition for trackable genetic variant libraries. Further provided herein are methods and compositions for recursive engineering. Further provided herein are methods and compositions for multiplex engineering. Further provided herein are methods and compositions for enriching for editing and trackable engineered sequences and cells using nucleic acid-guided nucleases.

Methods for generating barcoded combinatorial libraries

The present disclosure provides compositions and methods to increase the percentage of edited cells in a cell population when employing nucleic-acid guided editing, as well as automated multi-module instruments for performing these methods. Specifically, the disclosure relates to methods, compositions, modules and automated multi-module cell processing instruments that increase the efficiency of nucleic acid-guided editing in a cell population using a nucleic acid nuclease (i.e., an RNA-guided nuclease or 'RGN')/single-strand binding protein ('SSB') fusion system. The system leverages a single-strand binding protein (SSP) and single-strand DNA annealing protein (&#x201c;SSAP&#x201d;) interactions to drive enhanced recruitment.

Affinity tag for recombination protein recruitment

The present disclosure relates to methods and compositions that allow one to identify in vivo edited cells when employing nucleic-acid guided editing. Additionally provided are automated multi-module instruments for performing editing and selection methods and using the compositions.

Cascade/dCas3 complementation assays for in vivo detection of nucleic acid-guided nuclease edited cells

Described herein are synthetic oligonucleotides for editing a cell. The oligonucleotides described herein comprise the following covalently-linked components: (i) a nucleic acid encoding a guide RNA (gRNA) sequence targeting a target region in a cell; (ii) a region homologous to the target region comprising a change in sequence relative to the target region; and (iii) a site conferring immunity to nuclease-mediated editing.

Growing community of inventors

Boulder, CO, United States of America

Andrew Garst