公开(公告)号：US20230250420A1

公开(公告)日：2023-08-10

申请号：US18174397

申请日：2023-02-24

Applicant: Inscripta, Inc.

Inventor： Phillip Belgrader ,  Nathan Bade ,  Christian Siltanen ,  Aamir Mir ,  Xi-Jun Chen ,  Janine Mok ,  Burak Dura ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/10 ,  C12M1/34 ,  C12M1/26 ,  C12N15/113 ,  C12N15/86 ,  C12M3/00 ,  C12M3/04 ,  C12M1/00 ,  C12N5/074 ,  C12N15/11 ,  C12N15/88 ,  C12N15/90 ,  B01L3/00 ,  B01L7/00 ,  C12M1/32 ,  C12M3/06

CPC classification number: C12N15/1082 ,  C12M41/36 ,  C12M33/14 ,  C12N15/113 ,  C12N15/86 ,  C12M23/42 ,  C12M27/10 ,  C12M29/04 ,  C12M43/00 ,  C12M47/02 ,  C12N5/0696 ,  C12N15/1065 ,  C12N15/1068 ,  C12N15/11 ,  C12N15/88 ,  C12N15/907 ,  B01L3/502761 ,  B01L7/00 ,  C12M23/12 ,  C12M23/16 ,  C12M23/50 ,  C12M47/04 ,  C12N9/22

Abstract: Compositions of matter, methods, modules, and automated instruments may relate to synthesizing a library including an editing cassette including a different gRNA and donor DNA pair, amplifying the editing cassette in a partition separate from other editing cassettes in the library, adding nuclease to the partition, and adding lipofectamine to the editing cassette and nuclease to form a lipofectamine/nucleic acid/nuclease complex. A microcarrier coated in extracellular matrix or a cell adhesion molecule coating may be added to the lipofectamine/nucleic acid/nuclease complex. Cell growth material, the microcarrier, and mammalian cells may be transferred to a growth module in an automated closed cell editing instrument via a liquid handling system. The mammalian cells may be allowed to seed on the microcarrier. Conditions may be provided for the mammalian cells to take-up and be edited by a payload associated with the lipofectamine/nucleic acid/nuclease complex. The mammalian cells may be detached from the microcarrier.

公开(公告)号：US20220145327A1

公开(公告)日：2022-05-12

申请号：US17584298

申请日：2022-01-25

Applicant: Inscripta, Inc.

Inventor： Phillip Belgrader ,  Christian Siltanen ,  William Watterson ,  Burak Dura ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/86 ,  C12N9/22 ,  C12N15/11 ,  C12N5/074 ,  C12N5/071 ,  C12M1/36 ,  C12M1/12 ,  C12M1/06 ,  C12M1/02 ,  C12M1/34 ,  C12M1/00 ,  C12M1/26

Abstract: This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells.

公开(公告)号：US20220307014A1

公开(公告)日：2022-09-29

申请号：US17829263

申请日：2022-05-31

Applicant: Inscripta, Inc.

Inventor： Burak Dura ,  Phillip Belgrader ,  Christian Siltanen ,  William Watterson ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/10 ,  C12M1/34 ,  C12M1/26 ,  C12N15/113 ,  C12N15/86 ,  C12M3/00 ,  C12M3/04 ,  C12M1/00 ,  C12N5/074 ,  C12N15/11 ,  C12N15/88 ,  C12N15/90 ,  B01L3/00 ,  B01L7/00 ,  C12M1/32 ,  C12M3/06

Abstract: This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells.

公开(公告)号：US11591592B2

公开(公告)日：2023-02-28

申请号：US17239540

申请日：2021-04-23

Applicant: Inscripta, Inc.

Inventor： Phillip Belgrader ,  Nathan Bade ,  Christian Siltanen ,  Aamir Mir ,  Xi-Jun Chen ,  Janine Mok ,  Burak Dura ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/11 ,  C12N15/10 ,  C12M1/34 ,  C12M1/26 ,  C12N15/113 ,  C12N15/86 ,  C12M3/00 ,  C12M3/04 ,  C12M1/00 ,  C12N5/074 ,  C12N15/88 ,  C12N15/90 ,  B01L3/00 ,  B01L7/00 ,  C12M1/32 ,  C12M3/06 ,  C12N9/22

Abstract: This invention relates to compositions of matter, methods, modules and automated, end-to-end closed instruments for automated mammalian cell growth, reagent bundle creation and mammalian cell transfection followed by nucleic acid-guided nuclease editing in live mammalian cells. The disclosed compositions and method entail making “reagent bundles” comprising many (hundreds of thousands to millions) clonal copies of an editing cassette and delivering or co-localizing the reagent bundles with live mammalian cells such that the editing cassettes edit the cells and the edited cells continue to grow.

公开(公告)号：US11407994B2

公开(公告)日：2022-08-09

申请号：US17584298

申请日：2022-01-25

Applicant: Inscripta, Inc.

Inventor： Phillip Belgrader ,  Christian Siltanen ,  William Watterson ,  Burak Dura ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N5/00 ,  C12N15/63 ,  C12N15/85 ,  C07H21/04 ,  C12N15/10 ,  C12M1/34 ,  C12M1/26 ,  C12N15/113 ,  C12N15/86 ,  C12M3/00 ,  C12M3/04 ,  C12M1/00 ,  C12N5/074 ,  C12N15/11 ,  C12N15/88 ,  C12N15/90 ,  B01L3/00 ,  B01L7/00 ,  C12M1/32 ,  C12M3/06 ,  C12N9/22

Abstract: This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells. The present compositions and methods entail viral delivery of an editing cassette to live mammalian cells such that the editing cassettes edit the cells and the edited cells continue to grow, preferably using a fully-automated end-to-end instrument to process the cells without human intervention to enhance cell processing uniformity and to maintain the integrity of the cell culture.

公开(公告)号：US20210332389A1

公开(公告)日：2021-10-28

申请号：US17239540

申请日：2021-04-23

Applicant: Inscripta, Inc.

Inventor： Phillip Belgrader ,  Nathan Bade ,  Christian Siltanen ,  Aamir Mir ,  Xi-Jun Chen ,  Janine Mok ,  Burak Dura ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/90 ,  C12N15/10 ,  C12N5/074 ,  C12M1/00 ,  C12M3/00 ,  C12N15/88 ,  C12M3/04 ,  C12N15/11 ,  C12N9/22

Abstract: This invention relates to compositions of matter, methods, modules and automated, end-to-end closed instruments for automated mammalian cell growth, reagent bundle creation and mammalian cell transfection followed by nucleic acid-guided nuclease editing in live mammalian cells.

公开(公告)号：US20210332388A1

公开(公告)日：2021-10-28

申请号：US17237747

申请日：2021-04-22

Applicant: Inscripta, Inc.

Inventor： Phillip Belgrader ,  Christian Siltanen ,  Burak Dura ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/90 ,  C12N15/10 ,  C12M3/06 ,  C12M1/00 ,  C12M1/32 ,  B01L7/00 ,  C12N15/11 ,  C12N9/22 ,  B01L3/00

Abstract: This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth, reagent bundle creation and mammalian cell transfection followed by nucleic acid-guided nuclease editing in live mammalian cells.

公开(公告)号：US20210332352A1

公开(公告)日：2021-10-28

申请号：US17239538

申请日：2021-04-23

Applicant: Inscripta, Inc.

Inventor： Phillip Belgrader ,  Christian Siltanen ,  William Watterson ,  Burak Dura ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/10 ,  C12N15/113 ,  C12M1/26 ,  C12M1/34

Abstract: This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells.

公开(公告)号：US20240141331A1

公开(公告)日：2024-05-02

申请号：US18504405

申请日：2023-11-08

Applicant: Inscripta, Inc.

Inventor： Burak Dura ,  Phillip Belgrader ,  Christian Siltanen ,  William Watterson ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/10 ,  B01L3/00 ,  B01L7/00 ,  C12M1/00 ,  C12M1/26 ,  C12M1/32 ,  C12M1/34 ,  C12M3/00 ,  C12M3/04 ,  C12M3/06 ,  C12N5/074 ,  C12N15/11 ,  C12N15/113 ,  C12N15/86 ,  C12N15/88 ,  C12N15/90

CPC classification number: C12N15/1082 ,  B01L3/502761 ,  B01L7/00 ,  C12M23/12 ,  C12M23/16 ,  C12M23/42 ,  C12M23/50 ,  C12M27/10 ,  C12M29/04 ,  C12M33/14 ,  C12M41/36 ,  C12M43/00 ,  C12M47/02 ,  C12M47/04 ,  C12N5/0696 ,  C12N15/1065 ,  C12N15/1068 ,  C12N15/11 ,  C12N15/113 ,  C12N15/86 ,  C12N15/88 ,  C12N15/907 ,  C12N9/22

Abstract: This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells.

公开(公告)号：US11845932B2

公开(公告)日：2023-12-19

申请号：US17829263

申请日：2022-05-31

Applicant: Inscripta, Inc.

Inventor： Burak Dura ,  Phillip Belgrader ,  Christian Siltanen ,  William Watterson ,  Bruce Chabansky ,  David Stumbo ,  Eric Smith ,  Jorge Bernate

IPC: C12N15/63 ,  C12N5/00 ,  C12N15/79 ,  C07H21/04 ,  C12N15/10 ,  C12M1/34 ,  C12M1/26 ,  C12N15/113 ,  C12N15/86 ,  C12M3/00 ,  C12M3/04 ,  C12M1/00 ,  C12N5/074 ,  C12N15/11 ,  C12N15/88 ,  C12N15/90 ,  B01L3/00 ,  B01L7/00 ,  C12M1/32 ,  C12M3/06 ,  C12N9/22

CPC classification number: C12N15/1082 ,  B01L3/502761 ,  B01L7/00 ,  C12M23/12 ,  C12M23/16 ,  C12M23/42 ,  C12M23/50 ,  C12M27/10 ,  C12M29/04 ,  C12M33/14 ,  C12M41/36 ,  C12M43/00 ,  C12M47/02 ,  C12M47/04 ,  C12N5/0696 ,  C12N15/1065 ,  C12N15/1068 ,  C12N15/11 ,  C12N15/113 ,  C12N15/86 ,  C12N15/88 ,  C12N15/907 ,  B01L2200/0647 ,  B01L2300/0681 ,  B01L2300/123 ,  B01L2300/161 ,  B01L2400/0415 ,  B01L2400/0421 ,  B01L2400/0424 ,  C12N9/22 ,  C12N2310/20 ,  C12N2510/00 ,  C12N2740/10011 ,  C12N2740/15011 ,  C12N2750/14111 ,  C12N2800/80

Abstract: Nucleic acid-guided nuclease editing in mammalian cells may include passaging mammalian cells, in an automated closed cell editing instrument, into smaller aggregates when the aggregates exceed 50-300 microns in size. A library of viral particles may be delivered to the mammalian cells at a multiplicity of infection such that each mammalian cell receives one or no viral particle. The library may include viral vectors with an editing cassette including a pair of gRNA coding sequence and donor DNA. Conditions may be provided to allow a viral vector of the viral vectors to integrate into the mammalian cells. Enriching for mammalian cells may be done with an integrated viral vector. A nucleic acid-guided nuclease or nuclease fusion or a coding sequence for a nucleic acid-guided nuclease or nuclease fusion may be delivered to the enriched mammalian cells and conditions may be provided to allow editing in the mammalian cells.