Harvard College

Massachusetts Institute of Technology

The Invention Science Fund I LLC

The Borad Institute, Inc.

Kevin Michael Esvelt

David R Liu

George M Church

Prashant G Mali

Jacob Charles Carlson

Lowell L Wood, Jr

Clarence T Tegreene

Edward K Y Jung

Jordin T Kare

Eric Leuthardt

Philip Andrew Eckhoff

Mahalaxmi Gita Bangera

Erez Lieberman

Ahmed Hussein Badran

Brett M Bethke

Christopher Somogyi

Xiaoyan Robert Bao

Anna Bershteyn

Kyle B Gustafson

William Michael Kaminsky

Lily Yvonne Kim

Ankur Moitra

Jeremiah James Zartman

Dario G Amodei

Stephanie Yaung

Sarah Anna Vitak

Andrea L Smidler

Elizabeth Ashton Strait

Mariah Avila

Erika Alden DeBenedictis

Emma Chory

The invention relates, in part, to methods and systems with which to identify guide RNAs (gRNAs) and methods and systems with which to prepare, design, and generate gRNAs and minimally repetitive arrays of gRNAs.

Methods of multiplexing CRISPR

The invention relates, in part, to devices, systems and methods to increase the likelihood of survival and reproductive success of preselected organisms. An organism container may be configured for permitting access to a preselected organism and denying access to other organisms. The container may include a floor, one or more walls, a door, a lock associated with the door, and a first sensor configured to unlock the lock to selectively move the door into the open configuration when it detects the presence of the preselected organism at the door and outside of the organism enclosure so the preselected organism can enter the container cavity, and the first sensor is configured to prevent access to the container cavity to other organisms.

Devices, systems, and methods for introducing organisms into populations

Strategies, systems, methods, reagents, and kits for phage-assisted continuous evolution are provided herein. These include strategies, systems, methods, reagents, and kits allowing for stringency modulation to evolve weakly active or inactive biomolecule variants, negative selection of undesired properties, and/or positive selection of desired properties.

Negative selection and stringency modulation in continuous evolution systems

The invention provides systems, methods, reagents, apparatuses, vectors, and host cells for the continuous evolution of nucleic acids. For example, a lagoon is provided in which a population of viral vectors comprising a gene of interest replicates in a stream of host cells, wherein the viral vectors lack a gene encoding a protein required for the generation of infectious viral particles, and wherein that gene is expressed in the host cells under the control of a conditional promoter, the activity of which depends on a function of the gene of interest to be evolved. Some aspects of this invention provide evolved products obtained from continuous evolution procedures described herein. Kits containing materials for continuous evolution are also provided.

Continuous directed evolution

The invention, in part, includes compositions comprising quadruplet decoding tRNAs and their encoding sequences. The invention also includes assay methods to assess quadruplet decoding as well as methods of preparing quadruplet decoding suppression tRNAs.

Frameshift suppressor tRNA compositions and methods of use

Methods of modulating expression of a target nucleic acid in a cell are provided including introducing into the cell a first foreign nucleic acid encoding one or more RNAs complementary to DNA, wherein the DNA includes the target nucleic acid, introducing into the cell a second foreign nucleic acid encoding a nuclease-null Cas9 protein that binds to the DNA and is guided by the one or more RNAs, introducing into the cell a third foreign nucleic acid encoding a transcriptional regulator protein or domain, wherein the one or more RNAs, the nuclease-null Cas9 protein, and the transcriptional regulator protein or domain are expressed, wherein the one or more RNAs, the nuclease-null Cas9 protein and the transcriptional regulator protein or domain co-localize to the DNA and wherein the transcriptional regulator protein or domain regulates expression of the target nucleic acid.

RNA-guided transcriptional regulation

Methods of modulating expression of a target nucleic acid in a cell are provided including use of multiple orthogonal Cas9 proteins to simultaneously and independently regulate corresponding genes or simultaneously and independently edit corresponding genes.

Orthogonal Cas9 proteins for RNA-guided gene regulation and editing

Stably immunized cells and methods of making stably immunized cells are provided. Methods of altering the microbiota of an ecological environment are provided. Methods of modifying target chromosomes are provided. Methods of delivering genetic material to target cells are provided.

Precise microbiota engineering at the cellular level

RNA guided Cas9 gene drives and method for their use are disclosed.

RNA-guided gene drives

Growing community of inventors

Cambridge, MA, United States of America

Kevin Michael Esvelt