Automated cell processing methods, modules, instruments, and systems
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.
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 a protein.
28. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein a nuclease is delivered to the mammalian cells as an expression sequence for a protein.
29. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the nuclease is a MAD7 nuclease.
30. The automated stand-alone multi-module cell editing instrument of claim 13 , wherein the nuclease is a Cas9 nuclease.