Curing for recursive nucleic acid-guided cell editing

Abstract

The present disclosure provides automated multi-module instrumentation and automated methods for performing recursive editing of live cells with curing of editing vectors from prior rounds of editing.

1 . A composition of matter for recursive nucleic acid-directed nuclease CRISPR editing comprising:
 a first vector comprising:
 an origin of replication; 
 one or more editing gRNA and donor DNA pairs under the control of a first inducible promoter, wherein the editing gRNA and donor DNA pairs edit target sequences in a cell; 
 a curing target sequence, wherein the curing target sequence is cut to cleave the vector; 
 a first selectable marker; and 
   a second vector comprising:
 a second selectable marker; and 
 a curing gRNA sequence under the control of a second inducible promoter, wherein the curing gRNA targets the curing target sequence on the first vector. 
   
     
     
         2 .- 25 . (canceled)