System Biosciences, LLC

Leland Stanford Junior University

Fangting Wu

Kevin Kwei

Travis John Antes

Stefanie Morgan

Kevin Chun-Kai Wang

Chiao-Chain Huang

Chromatin looping is key to gene regulation, yet no broadly applicable methods to selectively modify chromatin loops have been described. Disclosed herein is an engineered method for chromatin loop reorganization using CRISPR-dCas9 (CLOuD9) to selectively and reversibly establish chromatin loops. Disclosed herein is the power of this technology to selectively modulate gene expression at targeted loci.

Manipulation of nuclear architecture

The invention relates to engineered CRISPR/Cas9 systems for genomic modification in mammalian cells. The present specification describes the design and testing of a polynucleotide encoding the() Cas9 protein, where the nucleotide sequence has been optimized for expression in mammalian cells. The specification also describes all-in-one systems for RNA-guided genome engineering in mammalian cells, including human cells.

Compositions and methods directed to CRISPR/Cas genomic engineering systems

The invention relates to engineered CRISPR/Cas9 systems for genomic modification and regulation of gene expression in mammalian cells. The specification describes the design and validation of polynucleotides encoding the() Cas9 gene and protein and variants of that protein, where the nucleotide sequence has been optimized for expression in mammalian cells, and also modified by fused sequences that enhance various aspects of the CRISPR/Cas system. The specification also describes systems for RNA-guided genome engineering and gene regulation in mammalian cells, including human cells.

CRISPR/Cas systems for genomic modification and gene modulation

The invention relates to compositions and methods for isolation of microvesicles produced by mammalian cells. These microvesicles, known as extracellular microvesicles or circulating microvesicles, are isolated from sample materials such as body fluids, or from cell culture media that has been used to culture and maintain mammalian cells in vitro. The isolation of microvesicles as described herein results in purification and concentration of the microvesicles. The invention also provides related methods for producing blood serum and/or blood plasma that is free of detectable microvesicles, largely depleted of microvesicles, or has reduced concentration of microvesicles compared to the blood serum or blood plasma starting material (collectively termed 'microvesicle-depleted'). The generation of microvesicle-depleted blood serum or plasma is critical for use in experimental systems where it is desirable to use a cell culture media that does not contain endogenous microvesicles, or has been depleted of endogenous microvesicles, from the source material.

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Mountain View, CA, United States of America

Fangting Wu