CELL LIBRARIES CREATED USING RATIONALLY DESIGNED NUCLEIC ACIDS
In an illustrative embodiment, cell libraries created by rationally designed nucleic acids comprising sequences to facilitate nuclease-directed genome editing events are provided. The cell libraries include saturation mutagenesis cell libraries, promoter swap cell libraries, SNP swap cell libraries, terminator swap cell libraries, and combinations thereof.
1 . A cell library created using an automated editing instrument for nuclease-directed genome editing, wherein the instrument comprises:
a housing;
a receptacle configured to receive cells and one or more rationally designed nucleic acids comprising sequences to facilitate nuclease-directed genome editing events in the cells;
a transformation unit for introduction of the nucleic acid(s) into the cells;
an editing unit for allowing the nuclease-directed genome editing events to occur in the cells, and
a processor-based system configured to operate the instrument based on user input, wherein the nuclease-directed genome editing events created by the automated instrument result in a cell library comprising individual cells with rationally designed edits.
2 . The cell library of claim 1 , wherein the nuclease-directed genome editing events in the cells creates a saturation mutagenesis cell library.
3 . The cell library of claim 1 , wherein the nuclease-directed genome editing events in the cells creates a promoter swap cell library.
4 . The cell library of claim 1 , wherein the nuclease-directed genome editing events in the cells creates a terminator swap cell library.
5 . The cell library of claim 1 , wherein the nuclease-directed genome editing events in the cells creates a SNP swap cell library.
6 . The cell library of claim 1 , wherein the nuclease-directed genome editing events in the cells creates a promoter and terminator swap cell library.
7 . The cell library of claim 1 , wherein the library comprises at least 100,000 edited cells.
8 . The cell library of claim 7 , wherein the library comprises at least 1,000,000 edited cells.
9 . The cell library of claim 1 , wherein the nuclease-directed genome editing is CRISPR-directed genome editing.
10 . A cell library created using an automated editing instrument for nuclease-directed genome editing, wherein the instrument comprises:
a housing;
a cell receptacle configured to receive cells;
a nucleic acid receptacle configured to receive one or more rationally designed nucleic acids comprising sequences to facilitate nuclease-directed genome editing events in the cells;
a transformation unit for introduction of the nucleic acid(s) into the cells;
an editing unit for allowing the nuclease-directed genome editing events to occur in the cells, and
a processor-based system configured to operate the instrument based on user input, wherein the nuclease-directed genome editing events created by the automated instrument result in a cell library comprising individual cells with rationally designed edits.
11 . The cell library of claim 10 , wherein the nuclease-directed genome editing events in the cells creates a saturation mutagenesis cell library.
12 . The cell library of claim 10 , wherein the nuclease-directed genome editing events in the cells creates a promoter swap cell library.
13 . The cell library of claim 10 , wherein the nuclease-directed genome editing events in the cells creates a terminator swap cell library.
14 . The cell library of claim 10 , wherein the nuclease-directed genome editing events in the cells creates a SNP swap cell library.
15 . The cell library of claim 10 , wherein the nuclease-directed genome editing events in the cells creates a promoter swap cell library.
16 . The cell library of claim 10 , wherein the library comprises at least 100,000 edited cells.
17 . The cell library of claim 16 , wherein the library comprises at least 1,000,000 edited cells.
18 . The cell library of claim 1 , wherein the nuclease-directed genome editing is CRISPR-directed genome editing.
19 . A cell library created using an automated editing instrument for recursive nuclease-directed genome editing, wherein the instrument comprises:
a housing;
means to receive cells and one or more rationally designed nucleic acids comprising sequences to facilitate nuclease-directed genome editing in the cells;
means for introduction of the nucleic acid(s) into the cells;
means for allowing the nuclease-directed genome editing events to occur,
means for the growth of the edited cells;
means for concentrating the edited cells;
means for collecting the edited cells; and
means for configuring the operation of the instrument based on user input,
wherein the nuclease-directed genome editing events are repeated two or more times within the automated instrument to create a cell library comprising individual cells with two or more rationally designed edits.
20 . The cell library of claim 19 , wherein the automated s instrument used to create the cell library further comprises means for selecting the edited cells.