Automated cell processing methods, modules, instruments, and systems

Patent metadata

Field	Value
Publication number	US-11293021-B1
Application number	US-202117538922-A
Country	US
Kind code	B1
Filing date	Nov 30, 2021
Priority date	Jun 23, 2016
Publication date	Apr 5, 2022
Grant date	Apr 5, 2022

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 mammalian cells;
one or more receptacles configured to receive expression vectors, wherein the expression vectors introduce a desired DNA change to the target region and remove a proto-spacer motif (PAM) region from the target region;
a growth module for growing the mammalian cells for transformation;
a transformation module configured to introduce the expression vectors into the mammalian cells;
a nuclease-directed editing module configured to allow the introduced expression vectors to edit nucleic acids in the mammalian cells;
a magnetic separation module configured to separate edited mammalian cells from unedited mammalian 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; from the one or more receptacles configured to receive expression vectors to the transformation module; from the transformation module to the nuclease-directed editing module; and from the nuclease-directed editing module to the magnetic separation module without user intervention.

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

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

4. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the automated stand-alone multi-module cell editing instrument further comprises a recovery module separate from the nuclease-directed editing module.

5. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the growth module is separate from the transformation module.

6. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the transformation module performs a process selected from electroporation, lipofection, optoporation, injection, microprecipitation, magnetofection, microinjection, particle bombardment, sonoporation, laser-induced poration, bead transfection, calcium phosphate and calcium chloride co-precipitation, and DEAE-dextran-mediated transfection.

7. The automated stand-alone multi-module cell editing instrument of claim 6 , wherein the transformation module performs electroporation.

8. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the transformation module is combined with the nuclease-directed editing module.

9. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the transformation module is separate from the nuclease-directed editing module.

10. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the growth module is combined with the transformation module.

11. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the growth module is separate from the transformation module.

12. The automated stand-alone multi-module cell editing instrument of claim 1 , wherein the growth module, transformation module and nuclease-directed editing module are combined.

13. An automated stand-alone multi-module cell editing instrument comprising:
a housing configured to house all or some of the modules;
one or more receptacles configured to receive expression vectors, wherein the expression vectors introduce a desired DNA change to the target region and remove a proto-spacer motif (PAM) region from the target region;
a growth module for growing mammalian cells for transformation;
a transformation module configured to introduce the expression vectors into the mammalian cells;
a nuclease-directed editing module configured to allow the introduced expression vectors to edit nucleic acids in the mammalian cells;
a magnetic separation module configured to separate edited mammalian cells from unedited mammalian 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 growth module to the transformation module; from the one or more receptacles configured to receive expression vectors to the transformation module; from the transformation module to the nuclease-directed editing module; and from the nuclease-directed editing module to the magnetic separation module without user intervention.

14. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the nuclease-directed editing module is also a recovery module.

15. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the automated stand-alone multi-module cell editing instrument further comprises a recovery module separate from the nuclease-directed editing module.

16. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the transformation module performs a process selected from electroporation, lipofection, optoporation, injection, microprecipitation, magnetofection, microinjection, particle bombardment, sonoporation, laser-induced poration bead transfection, calcium phosphate and calcium chloride co-precipitation, and DEAE-dextran-mediated transfection.

17. The automated stand-alone multi-module cell editing instrument of claim 16 , wherein the transformation module performs bead transfection.

18. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the transformation module is combined with the nuclease-directed editing module.

19. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the transformation module is separate from the nuclease-directed editing module.

20. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the growth module is combined with the transformation module.

21. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the growth module is separate from the transformation module.

22. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the growth module, transformation module and nuclease-directed editing module are combined.

23. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the expression vectors are viral vectors.

24. The automated stand-alone multi-module cell editing instrument of claim 23 , wherein the viral vector is selected from a retrovirus vector, replication defective retrovirus vector, adenovirus vector, replication defective adenovirus vector, and adeno-associated virus vector and a lentivirus vector.

25. The automated stand-alone multi-module cell editing instrument of claim 24 , wherein the viral vector is an adenovirus vector.

26. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the mammalian cells are human cells.

27. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein a nuclease is delivered to the mammalian cells as

Assignees

• Inscripta Inc

Inventors

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

Classifications

• C12N15/1082 Primary

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

• C12M41/48 Primary

  Automatic or computerized control (automatic analysis G01N35/00) · CPC title

• C12N2310/20

  involving clustered regularly interspaced short palindromic repeats [CRISPR] · CPC title

• C12M37/04

  Seals · CPC title

• C12M23/44

  Multiple separable units; Modules · CPC title