Leland Stanford Junior University

The Brigham and Women's Hospital, Inc.

Beth Israel Deaconess Medical Center, Inc.

Brandeis University

European Molecular Biology Laboratory

Lars Steinmetz

Ronald W Davis

Utkan Demirci

Naside Gozde Durmus

Murat Baday

Thiruppathiraja Chinnasamy

Savas Tasoglu

Ionita C Ghiran

Semih Calamak

Jaeyoung Yang

Alessandro Tocchio

Robert P St Onge

Huseyin Cumhur Tekin

James E Haber

Kevin Roy

Han Sun

Justin D Smith

Francesca Briganti

Wu Wei

Ionita Ghiran

A heterogeneous population of cells are separated and collected according to a method. The heterogeneous population of cells in a paramagnetic medium are placed in a fluidic channel in which the fluidic channel comprises two or more outlets. The heterogeneous population of cells in the fluidic channel are separated based on differences in magnetic susceptibility and density of the heterogeneous population of cells. Fluid comprising the separated cells is withdrawn from the two or more outlets using variable flow rates by fluidic pumps at respective ones of the two or more outlets simultaneously to fractionalize the fluid comprising the separated cells across the two or more outlets by manipulation of the variable flow rates relative to one another.

Sorting biological and non-biological moieties using magnetic levitation

The present disclosure relates to multiplex production and phenotyping of genetically engineered cells using RNA-guided nucleases and genomic barcoding. In particular, high-throughput multiplex genome editing is achieved utilizing a system that facilitates precise genome editing at desired target chromosomal loci by homology directed repair. Integration of guide RNA and donor DNA sequences as a genomic barcode at a separate chromosomal locus allows identification, isolation, and massively-parallel validation of individual variants from a pool of transformants. Strains can be arrayed according to their precise genetic modifications, as specified by donor DNA incorporation in heterologous or native genes. The present disclosure further relates to a method of editing codons outside of canonical guide RNA recognition regions, which enables complete saturation mutagenesis of protein-coding genes, a marker-based internal cloning method, which removes background due to oligonucleotide synthesis errors and incomplete vector backbone cleavage, and a method of enhancing homology directed repair by active donor recruitment.

Multiplex production and barcoding of genetically engineered cells

Methods of treatment, genetic screening, and disease models for heart conditions associated with RBM20 deficiency are disclosed. In particular, methods of treating heart conditions associated with RBM20 deficiency, including RBM20-dependent dilated cardiomyopathy and heart failure with compounds that upregulate expression of RBM20, such as all-trans retinoic acid are provided. Also disclosed are methods of genetic screening to detect the presence of a P633L mutation in RBM20 in order to identify individuals having a genetic predisposition to developing RBM20-dependent DCM. Induced pluripotent stem cell-derived cardiomyocytes (IPSC-CMs) produced by differentiation of IPSCs comprising at least one RBM20 allele encoding a P633L mutation and methods of using them in screening for therapeutics for treating RBM20-dependent DCM are also disclosed.

Methods of treatment, genetic screening, and disease models for heart conditions associated with RBM20 deficiency

Magnetic cell levitation and cell monitoring systems and methods are disclosed. A method for separating a heterogeneous population of cells is performed by placing a microcapillary channel containing the heterogeneous population of cells in a magnetically-responsive medium in the disclosed levitation system and separating the cells by balancing magnetic and corrected gravitational forces on the individual cells. A levitation system is also disclosed, having a microscope on which the microcapillary channel is placed and a set of two magnets between which the microcapillary channel is placed. Additionally, a method for monitoring cellular processes in real-time using the levitation system is disclosed.

System and method for cell levitation and monitoring

Systems and methods for levitating populations of moieties, cells, or other such units using one or more magnets in a microfluidic environment are provided. These systems and methods may be used to, for example, separate or sort heterogeneous populations of the units from one another, to assembly a multi-unit assembly during the levitating of the units, and to evaluate samples at the point of care in real-time. These systems and methods may also utilize a frame that enables an imaging device, such as a smartphone, to capture the units in real time as they are manipulated in the system.

Growing community of inventors

San Francisco, CA, United States of America

Lars Steinmetz