Automated cell processing methods, modules, instruments, and systems comprising flow-through electroporation devices

• US 10,508,288 B1
• Filed: 09/14/2019
• Issued: 12/17/2019
• Est. Priority Date: 09/30/2017
• Status: Active Grant

• Current Assignee: Inscripta, Inc.
• Original Assignee: Inscripta, Inc.
• Inventors: Bernate, Jorge, Masquelier, Don, Belgrader, Phillip, Ness, Kevin
• Primary Examiner(s): Bowers, Nathan A
• Application Number:
  US16/571,087
• Time in Patent Office: 94 Days
• CPC Class Codes: C12M 23/44 Multiple separable units; Modules
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/81 for yeasts
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

In an illustrative embodiment, automated multi-module cell editing instruments comprising one or more flow-through electroporation devices or modules are provided to automate genome editing in live cells.

Claims

1. An automated multi-module cell editing instrument comprising:

a housing configured to house all of some of the modules;

one or more receptacles configured to receive cells and nucleic acids;

a flow-through electroporation (FTEP) module configured to introduce the nucleic acids into the cells, wherein the FTEP module comprises;

a. at least a first inlet and a first inlet channel for introducing a fluid comprising the cells and the nucleic acids into the FTEP module;

b. an outlet and an outlet channel for removing a fluid comprising transformed cells from the FTEP module;

c. a flow channel intersecting and positioned between a first inlet channel and the outlet channel; and

d. two or more electrodes positioned in the flow channel between the intersection of the flow channel with the first inlet channel and the intersection of the flow channel with the outlet channel, wherein the electrodes apply one or more electric pulses to the cells in the fluid as they pass through the flow channel thereby introducing the nucleic acids into the cells in the fluid;

a selection module configured to select for transformed cells;

an editing module configured to allow the nucleic acids to edit nucleic acids in the transformed 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 one or more receptacles to the FTEP module, from the FTEP module to the selection module, and from the selection module to the editing module, all without user intervention.

2. The automated multi-module cell editing instrument of claim 1 wherein the selection module is also the editing module.

3. The automated multi-module cell editing instrument of claim 1 wherein the selection module is separate from the editing module.

4. The automated multi-module cell editing instrument of claim 1 wherein the FTEP module further comprises a second inlet and a second inlet channel.

5. The automated multi-module cell editing instrument of claim 4 wherein the second inlet and second inlet channel of the FTEP module are located between the first inlet and first inlet channel and the electrodes of the FTEP module.

6. The automated multi-module cell editing instrument of claim 4 wherein the second inlet and second inlet channel of the FTEP module are located between the electrodes and the outlet channel and outlet of the FTEP module.

7. The automated multi-module cell editing instrument of claim 1 wherein the flow channel is constricted between the first inlet and first inlet channel and the outlet and outlet channel.

8. The automated multi-module cell editing instrument of claim 1 wherein the FTEP module further comprises a filter between the first inlet channel and the electrodes.

9. The automated multi-module cell editing instrument of claim 1, wherein device is configured for use with bacterial, yeast and mammalian cells.

10. The automated multi-module cell editing instrument of claim 1, further comprising a reagent cartridge.

11. The automated multi-module cell editing instrument of claim 10, wherein the FTEP is part of the reagent cartridge.

12. An automated multi-module cell editing instrument comprising a flow-through electroporation (FTEP) module, wherein the multi-module cell editing instrument comprises:

a housing configured to house all of some of the modules;

one or more receptacles configured to receive nucleic acids and cells;

a growth module in which to grow the cells;

the FTEP module configured to introduce the nucleic acids into the cells;

wherein the FTEP module comprises;

a. at least a first inlet and at least a first inlet channel for introducing a fluid comprising cells and nucleic acids to the FTEP module;

b. an outlet and an outlet channel for removing transformed cells and exogenous material from the FTEP module;

c. a flow channel positioned between the first inlet channel and the outlet channel; and

d. an electrode positioned on either side of the flow channel, wherein the electrodes apply one or more electric pulses to the cells in the fluid as they pass through the flow channel, thereby introducing the nucleic acids into the cells in the fluid;

a selection module;

an 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 one or more receptacles to the growth module, from the growth module to the FTEP module, from the FTEP module to the selection module, from the selection module to the editing module, all without user intervention.

13. The automated multi-module cell editing instrument of claim 12 wherein the selection module comprises an antibiotic.

14. The automated multi-module cell editing instrument of claim 12 wherein the selection module is separate from the editing module.

15. The automated multi-module cell editing instrument of claim 12 wherein the selection module is combined with the editing module.

16. The automated multi-module cell editing instrument of claim 12 wherein the electrodes define a narrowed portion of the flow channel.

17. The automated multi-module cell editing instrument of claim 16 wherein the electrodes are in direct contact with the fluid in the flow channel.

18. The automated multi-module cell editing instrument of claim 12 wherein the FTEP module further comprises a filter between the at least first inlet channel and the electrodes.

19. The automated multi-module cell editing instrument of claim 12 further comprising a reagent cartridge.

20. The automated multi-module cell editing instrument of claim 19 wherein the FTEP is a component of the reagent cartridge.