Whitehead Institute for Biomedical Research

Massachusetts Institute of Technology

Sangamo Biosciences, Inc.

Washington University

Dana-farber-cancer Institute Inc.

Sangamo Therapeutics, Inc.

Rudolf Jaenisch

Konrad Hochedlinger

Ruth Foreman

Yaqub Hanna

Richard A Young

Alexander Marson

Alexander Meissner

G Grant Welstead

Brett Chevalier

Frank Soldner

Marius Wernig

Christopher J Lengner

Oliver Tobias Brambrink

Lei Zhang

Josee Laganiere

Bryce Woodbury Carey

Hao Wu

Xin Tang

Thorold W Theunissen

X Shawn Liu

Nathanael Schiander Gray

Fyodor Urnov

Michael C Holmes

Philip D Gregory

Nathan Richard Wilson

Joel Voldman

Mriganka Sur

Alison Skelley

Jacob Hanna

Shuyuan Yao

Daniela Tropea

Yun Li

Kevin C Eggan

Yonatan Stelzer

Shafqat Khan

Dirk Hockemeyer

Oktay Kirak

Julien Muffat

Emanuela Giacometti

The present disclosure provides methods and compositions for inducing, maintaining and/or passaging naïve pluripotent stem cell. In some embodiments, the methods are performed in the absence of MEK inhibition which has been shown to result in genomic instability of naïve pluripotent stem cells.

Enhanced methods for inducing and maintaining naive human pluripotent stem cells

The present disclosure provides compounds of any one of Formulae (A) to (L). The present disclosure also provides compositions, uses, and methods that include or involve a compound described herein, a serine/threonine-protein kinase B-Raf (BRAF) inhibitor, an epidermal growth factor receptor (EGFR) inhibitor, a vascular endothelial growth factor 1 (VEGFR1) inhibitor, a fibroblast growth factor receptor 1 (FGFR1) inhibitor, or a combination thereof. The compounds, compositions, uses, and methods are useful in changing the pluripotency state of a vertebrate cell to a more naïve state.

Uses of kinase inhibitors for inducing and maintaining pluripotency

The invention relates to methods of modifying DNA methylation by contacting a cell with a catalytically inactive site specific nuclease fused to an effector domain having methylation or demethylation activity and one or more guide sequences.

Methods of editing DNA methylation

Potassium chloride cotransporter-2 (KCC2) plays a critical role in brain function, and deficiency in KCC2 has been linked to neurological diseases, psychiatric disorders, and central nervous system injuries. In particular, Rett syndrome (RTT), a severe neurodevelopmental disorder caused by mutations in the X-linked gene Methyl CpG binding Protein 2 (MECP2), has been linked to deficits in KCC2. The disclosure reports the use of CRISPR/Cas9 genome-editing technology to generate stem cell-derived, genetically defined KCC2 reporter human neurons for large-scale compound screening. This screening platform has been utilized to identify a number of small molecule compounds that are capable of enhancing KCC2 expression in both wild-type and RTT neurons, as well as organotypical brain slices cultured from wild-type mice. These first-in class compounds may be applied as a novel therapeutic approach to restore the impaired balance between excitation and inhibition observed in neurological diseases, psychiatric disorders, and central nervous system injuries.

KCC2 expression enhancing compounds and uses thereof

The invention provides methods for reprogramming somatic cells to generate multipotent or pluripotent cells. Such methods are useful for a variety 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 less differentiated state.

Methods for reprogramming somatic cells

The disclosure relates to a method of reprogramming one or more somatic cells, e.g., partially differentiated or fully/terminally differentiated somatic cells, to a less differentiated state, e.g., a pluripotent or multipotent state. In further embodiments the invention also relates to reprogrammed somatic cells produced by methods of the invention, to chimeric animals comprising reprogrammed somatic cells of the invention, to uses of said cells, and to methods for identifying agents useful for reprogramming somatic cells.

Nucleic acid constructs encoding reprogramming factors linked by self-cleaving peptides

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

The invention relates to methods of modifying DNA methylation by contacting a catalytically inactive site specific nuclease fused to an effector domain having methylation or demethylation activity and one or more guide sequences.

Described herein are cell culture media useful for the differentiation of human pluripotent stem cells into microglia. The methods described herein relate to in vitro generation of expandable, bankable, microglial cells by directed differentiation from human pluripotent stem cells (induced or embryonic). Using only defined cell culture media, differentiation of pluripotent stem cells is directed down a mesodermal path, in a rapid and scalable fashion, to generate cells adopting signatures of their in vivo counterparts, including gene expression, protein marker expression and functionality.

Culture medium for generating microglia from pluripotent stem cells and related methods

The disclosure relates to a method of reprogramming one or more somatic cells, e.g., partially differentiated or fully/terminally differentiated somatic cells, to a less differentiated state, e.g., a pluripotent or multipotent state. In further embodiments the invention also relates to reprogrammed somatic cells produced by methods of the invention, to uses of said cells, and to methods for identifying agents useful for reprogramming somatic cells.

Reprogramming of somatic cells

In some aspects, described herein is a DNA methylation reporter. In some aspects, the DNA methylation reporter comprises a promoter whose activity can be affected by exogenous methylation changes without being independently regulated by the DNA methylation machinery, operably linked to a DNA sequence that encodes a reporter molecule. In some embodiments the DNA methylation reporter comprises (i) a promoter derived from a mammalian imprinted gene promoter; and (ii) a sequence that encodes a reporter molecule that is detectable in individual mammalian cells, wherein the promoter is operably linked to the sequence that encodes the reporter molecule. Also described are nucleic acids that comprise the DNA methylation reporter, cells that have the DNA methylation reporter integrated into their genome, and non-human mammals comprising cells that have the DNA methylation reporter integrated into their genome. Also described are methods of measuring DNA methylation of a region of interest located in proximity to the DNA methylation reporter in the genome of a cell by detecting the reporter molecule.

Reporter of genomic methylation and uses thereof

Growing community of inventors

Brookline, MA, United States of America

Rudolf Jaenisch