公开(公告)号：US11103870B2

公开(公告)日：2021-08-31

申请号：US16259538

申请日：2019-01-28

申请人： Berkeley Lights, Inc.

发明人： Gregory G. Lavieu ,  Annamaria Mocciaro ,  Xiao Guan Radstrom ,  Jason M. McEwen ,  Magali Soumillon ,  J. Tanner Nevill ,  Volker L. S. Kurz ,  Patricia A. Dyck ,  Ravi K. Ramenani

IPC分类号： G01N15/10 ,  B01L3/00 ,  C12M3/06 ,  C12N15/10 ,  C12N15/00 ,  G01N15/02

摘要： Methods are described herein for isolating clonal populations of cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the individual cells into respective clonal populations of cells; and detecting, in one or more cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of cells. Also described are methods of performing genome editing within a microfluidic device, and compositions comprising one or more clonal populations of cells generated according to the methods disclosed herein.

公开(公告)号：US10239058B2

公开(公告)日：2019-03-26

申请号：US15802174

申请日：2017-11-02

申请人： Berkeley Lights, Inc.

发明人： Gregory G. Lavieu ,  Annamaria Mocciaro ,  Xiao Guan Radstrom ,  Jason M. McEwen ,  Magali Soumillon ,  J. Tanner Nevill ,  Volker L. S. Kurz ,  Patricia A. Dyck ,  Ravi K. Ramenani

IPC分类号： G01N15/00 ,  B01L3/00 ,  C12M3/06 ,  C12N15/10 ,  G01N15/10 ,  B03C7/02 ,  G01N15/02

摘要： Methods are described herein for isolating clonal populations of cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the individual cells into respective clonal populations of cells; and detecting, in one or more cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of cells. Also described are methods of performing genome editing within a microfluidic device, and compositions comprising one or more clonal populations of cells generated according to the methods disclosed herein.

公开(公告)号：US20190217297A1

公开(公告)日：2019-07-18

申请号：US16259538

申请日：2019-01-28

申请人： Berkeley Lights, Inc.

发明人： Gregory G. Lavieu ,  Annamaria Mocciaro ,  Xiao Guan Radstrom ,  Jason M. McEwen ,  Magali Soumillon ,  J. Tanner Nevill ,  Volker L.S. Kurz ,  Patricia A. Dyck ,  Ravi K. Ramenani

IPC分类号： B01L3/00 ,  G01N15/10 ,  C12N15/10 ,  C12M3/06

CPC分类号： B01L3/502761 ,  B01L3/502792 ,  B01L2200/0647 ,  B01L2200/0668 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2400/0424 ,  B01L2400/0427 ,  C12M23/16 ,  C12N15/1003 ,  C12N15/1024 ,  C12N15/1086 ,  C12N2310/00 ,  C12N2510/00 ,  G01N15/10 ,  G01N2015/0288 ,  G01N2015/1081

摘要： Methods are described herein for isolating clonal populations of cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the individual cells into respective clonal populations of cells; and detecting, in one or more cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of cells. Also described are methods of performing genome editing within a microfluidic device, and compositions comprising one or more clonal populations of cells generated according to the methods disclosed herein.

公开(公告)号：US11639495B2

公开(公告)日：2023-05-02

申请号：US16455118

申请日：2019-06-27

申请人： The Regents of the University of California ,  Berkeley Lights, Inc.

发明人： Alexander Marson ,  Gregory G. Lavieu ,  Annamaria Mocciaro ,  Theodore L. Roth ,  Magali Soumillon ,  Hayley M. Bennett

IPC分类号： C12N5/0783 ,  B01L3/00 ,  C12N15/10

摘要： Methods are described herein for isolating clonal populations of T cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual T cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the T cells into respective clonal populations of T cells; detecting, in one or more T cells of each clonal population, the absence of a cell surface marker that was present in the individual T cells (or precursors thereof); and detecting, in one or more T cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of T cells. Also described are compositions comprising one or more clonal populations of T cells isolated according to the methods disclosed herein.

公开(公告)号：US20200171501A1

公开(公告)日：2020-06-04

申请号：US16661310

申请日：2019-10-23

申请人： Berkeley Lights, Inc.

发明人： Jason M. McEwen ,  Magali Soumillon ,  Shao Ning Pei ,  Randall D. Lowe, Jr. ,  Samira A. Nedungadi ,  Volker L.S. Kurz ,  Jian Gong ,  Yara X. Mejia Gonzalez ,  Mckenzi S. Toh ,  Brian A. Rabkin ,  Jason C. Briggs ,  Darcy K. Kelly-Greene ,  James M. Porter, Jr.

IPC分类号： B01L3/00 ,  B01L7/00 ,  B03C5/00 ,  B03C5/02

摘要： Microfluidic devices having an electrowetting configuration and an optimized droplet actuation surface are provided for processing biological cells, e.g., for use in nucleic acid library preparation and/or synthesis (including amplification). The devices include a dielectric layer, a hydrophobic layer covalently bonded to the dielectric layer, and a first electrode. Methods of nucleic acid library preparation and/or synthesis can involve providing reagents to cells or nucleic acids by merging appropriate droplets on a droplet actuation surface within a water-immiscible organic liquid and can be performed in the presence of appropriate surfactants. The hydrophobic layer features self-associating molecules covalently bonded to a surface of the dielectric layer in a manner that produces a densely-packed monolayer that resists intercalation and or penetration by polar molecules or species. Also provided are systems for temperature control of the microfluidic device during nucleic acid library preparation and/or synthesis which can reduce temperature overshooting during heating and cooling steps.

公开(公告)号：US20200048606A1

公开(公告)日：2020-02-13

申请号：US16455118

申请日：2019-06-27

申请人： The Regents of the University of California ,  Berkeley Lights, Inc.

发明人： Alexander Marson ,  Gregory G. Lavieu ,  Annamaria Mocciaro ,  Theodore L. Roth ,  Magali Soumillon ,  Hayley M. Bennett

IPC分类号： C12N5/0783 ,  B01L3/00 ,  C12N15/10

摘要： Methods are described herein for isolating clonal populations of T cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual T cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the T cells into respective clonal populations of T cells; detecting, in one or more T cells of each clonal population, the absence of a cell surface marker that was present in the individual T cells (or precursors thereof); and detecting, in one or more T cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of T cells. Also described are compositions comprising one or more clonal populations of T cells isolated according to the methods disclosed herein.

公开(公告)号：US20180147576A1

公开(公告)日：2018-05-31

申请号：US15802174

申请日：2017-11-02

申请人： Berkeley Lights, Inc.

发明人： Gregory G. Lavieu ,  Annamaria Mocciaro ,  Xiao Guan Radstrom ,  Jason M. McEwen ,  Magali Soumillon ,  J. Tanner Nevill ,  Volker L.S. Kurz ,  Patricia A. Dyck ,  Ravi K. Ramenani

IPC分类号： B01L3/00 ,  C12M3/06 ,  C12N15/10 ,  G01N15/10

CPC分类号： B01L3/502761 ,  B01L3/502792 ,  B01L2200/0647 ,  B01L2200/0668 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2400/0424 ,  B01L2400/0427 ,  C12M23/16 ,  C12N15/1003 ,  C12N15/1024 ,  C12N15/1086 ,  C12N2310/00 ,  C12N2510/00 ,  G01N15/10 ,  G01N2015/0288 ,  G01N2015/1081

摘要： Methods are described herein for isolating clonal populations of cells having a defined genetic modification. The methods are performed, at least in part, in a microfluidic device comprising one or more sequestration pens. The methods include the steps of: maintaining individual cells (or precursors thereof) that have undergone a genomic editing process in corresponding sequestration pens of a microfluidic device; expanding the individual cells into respective clonal populations of cells; and detecting, in one or more cells of each clonal population, the presence of a first nucleic acid sequence that is indicative of the presence of an on-target genome edit in the clonal population of cells. Also described are methods of performing genome editing within a microfluidic device, and compositions comprising one or more clonal populations of cells generated according to the methods disclosed herein.