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US 2022/0033855 A1Published application
Arrayed nucleic acid-guided nuclease or nickase fusion editing
AssigneeInscripta, Inc.
InventorsAndrew Garst, Christian Siltanen
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Abstract
The present disclosure relates to methods for performing arrayed nucleic acid-guided nuclease nickase fusion editing allowing for rapid genotypic/phenotypic correlation without sequencing.
Claims30
A method for editing a population of live cells with a library of editing vectors comprising rationally-designed editing cassettes in situ comprising:
- designing and synthesizing a library of editing cassettes on a substrate wherein each editing cassette comprises a gRNA and a repair template and wherein each different editing cassette is in a different partition;
- washing in first single-stranded supplemental oligonucleotides encoding at least one promoter and at least one first primer site and at least one region complementary to the editing cassettes;
- performing PCR in the partitions to produce amplified editing cassettes;
- releasing the amplified editing cassettes from the substrate in the partition;
- adding cells to the partition;
- adding transformation reagents to each partition;
- transforming the cells with the amplified editing cassettes to produce transformed cells;
- allowing editing to take place in the transformed cells to produce edited cells;
- making a replica of the substrate;
- and
- phenotyping the edited cells.
8 more elements
A method for editing a population of live cells with a library of editing vectors comprising rationally-designed editing cassettes in situ comprising:
- designing and synthesizing a library of editing cassettes on a substrate wherein each editing cassette comprises a gRNA and a repair template and wherein each different editing cassette is in a different partition;
- washing in first single-stranded supplemental oligonucleotides encoding at least one promoter and at least one first primer site and at least one region complementary to the editing cassettes;
- releasing the amplified editing cassettes from the substrate in the partition;
- performing PCR in the partitions to produce amplified editing cassettes;
- adding cells to the partition;
- adding transformation reagents to each partition;
- transforming the cells with the amplified editing cassettes to produce transformed cells;
- allowing editing to take place in the transformed cells to produce edited cells;
- making a replica of the substrate;
- and
- phenotyping the edited cells.
8 more elements
Dependencies are inferred from the claim text.