Automated cell processing methods, modules, instruments, and systems

Patent metadata

Field	Value
Publication number	US-10947532-B2
Application number	US-201916412195-A
Country	US
Kind code	B2
Filing date	May 14, 2019
Priority date	Jun 30, 2017
Publication date	Mar 16, 2021
Grant date	Mar 16, 2021

Abstract

In an illustrative embodiment, automated multi-module cell editing instruments are provided to automate multiple edits into nucleic acid sequences inside one or more cells.

First claim

We claim:

1. An automated stand-alone multi-module cell editing instrument comprising:
a housing configured to house all or some of the modules;
a receptacle configured to receive cells;
one or more receptacles configured to receive nucleic acids;
a growth module for growing cells for transformation;
a transformation module configured to introduce the nucleic acids into the cells, wherein the transformation module comprises a single-unit electroporation device comprising a housing, an engagement member configured to engage with a pipette, a filter, an electroporation cuvette, and electrodes;
a nuclease-directed editing module configured to allow the introduced nucleic acids to edit nucleic acids in the cells;
a processor configured to operate the automated multi-module cell editing instrument based on user input and/or selection of a pre-programmed script; and
an automated liquid handling system to move liquids from the cell receptacle to the growth module, from the growth module to the transformation module and from the transformation module to the nuclease-directed editing module without user intervention.

2. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the nucleic acids in the one or more receptacles comprise a backbone and an editing cassette, and the automated multi-module cell editing instrument further comprises a nucleic acid assembly module.

3. The automated stand-alone multi-module cell editing instrument of claim 2 , wherein the nucleic acid assembly module is configured to perform isothermal nucleic acid assembly.

4. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the automated liquid handling system comprises a sipper or pipettor.

5. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the editing module is also a recovery module.

6. The automated stand-alone multi-module cell editing instrument of claim 1 , further comprising a recovery module separate from the editing module.

7. The automated stand-alone multi-module cell editing instrument of claim 1 , further comprising a selection module.

8. The automated stand-alone multi-module cell editing instrument of claim 7 , wherein the selection module is a same module as the nuclease-directed editing module.

9. The automated stand-alone multi-module cell editing instrument of claim 7 , wherein the selection module is separate from the nuclease-directed editing module.

10. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the receptacle configured to receive cells and the one or more receptacles configured to receive nucleic acids are configured to be contained within a reagent cartridge.

11. The automated stand-alone multi-module cell editing instrument of claim 10 , wherein some or all reagents required for cell editing are received by the reagent cartridge.

12. The automated stand-alone multi-module cell editing instrument of claim 11 , wherein the reagents contained within the reagent cartridge are locatable by a script read by the processor.

13. The automated stand-alone multi-module cell editing instrument of claim 12 , wherein the reagent cartridge includes reagents and is provided in a kit.

14. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the growth module comprises a rotating growth vial.

15. The automated stand-alone multi-module cell editing instrument of claim 14 , wherein the rotating growth vial comprises paddles.

16. The automated stand-alone multi-module cell editing instrument of claim 1 , further comprising a filtration module configured to concentrate the cells and render the cells electrocompetent.

17. An automated stand-alone multi-module cell editing instrument comprising:
a housing configured to house some or all of the modules;
a receptacle configured to receive cells;
at least one receptacle configured to receive nucleic acids for desired genome editing events in the cells;
a nucleic acid assembly module configured to assemble a vector backbone and an editing cassette, wherein the nucleic acid assembly module is configured to accept and assemble nucleic acids to facilitate the desired genome editing events in the cells;
a growth module configured to grow the cells;
a transformation module to introduce assembled nucleic acids into the cells, wherein the transformation module comprises a single-unit electroporation device comprising a housing, an engagement member configured to engage with a pipette, a filter, an electroporation cuvette, and electrodes;
a nuclease-directed editing module configured to allow the assembled nucleic acids to edit nucleic acids in the cells;
a processor configured to operate the automated multi-module cell editing instrument based on user input and/or selection of a script; and
an automated liquid handling system to move liquids from the cell receptacle to the growth module, from the growth module to the transformation module, and from the transformation module to the nuclease-directed editing module, as well as from the nucleic acid receptacle to the nucleic acid assembly module, and from the nucleic acid assembly module to the transformation module, all without user intervention.

18. The automated stand-alone multi-module cell editing instrument of claim 17 , further comprising at least one reagent cartridge containing reagents to perform cell editing in the automated multi-module cell editing instrument.

19. The automated stand-alone multi-module cell editing instrument of claim 18 , wherein the receptacles for the cells and nucleic acids are disposed within the reagent cartridge.

20. An automated stand-alone multi-module cell editing instrument comprising:
a housing configured to house some or all of the modules;
a growth module configured to grow cells;
a filtration module configured to concentrate the cells and render the cells electrocompetent;
a transformation module, wherein the transformation module comprises a single-unit electroporation device comprising a housing, an engagement member configured to engage with a pipette, a filter, an electroporation cuvette, and electrodes;
a combination recovery and nuclease-directed editing module configured to allow the cells to recover after electroporation in the transformation module and to allow the nucleic acids to edit the cells;
a processor configured to operate the automated multi-module cell editing instrument based on user input; and
an automated liquid handling system to move liquids from the growth module to the filtration module, from the filtration module to the transformation module, and from the transformation module to the recovery and nuclease-directed editing module, without user intervention.

Assignees

• Inscripta Inc

Inventors

• Masquelier Don
• Belgrader Phillip
• Bernate Jorge
• Gill Ryan
• Ness Kevin

Classifications

• C12M35/02

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12Q1/6865

  Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS] · CPC title

• C12N15/1082 Primary

  Preparation or screening gene libraries by chromosomal integration of polynucleotide sequences, HR-, site-specific-recombination, transposons, viral vectors · CPC title

• C12M41/36

  of biomass, e.g. colony counters or by turbidity measurements (electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404) · CPC title

• C12M37/04

  Seals · CPC title