Methods and compositions for genome editing in non-dividing cells
AssigneeThe Salk Institute for Biological Studies
InventorsJuan Carlos Izpisua Belmonte, Keiichiro Suzuki, Reyna Hernandez-Benitez, Jun Wu, Yuji Tsunekawa
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Abstract
Disclosed herein are homology-independent targeted integration methods of integrating an exogenous DNA sequence into a genome of a non-dividing cell and compositions for such methods. Methods herein comprise contacting the non-dividing cell with a composition comprising a targeting construct comprising the exogenous DNA sequence and a targeting sequence, a complementary strand oligonucleotide homologous to the targeting sequence, and a nuclease, thereby altering the genome of the non-dividing cell.
Claims20
Recombinant vectors configured to integrate an exogenous DNA sequence into a target nucleic acid sequence in a genome of a cell, wherein the vectors collectively encode:
- a targeting construct comprising the exogenous DNA sequence and a target sequence entirely in reverse orientation with respect to the exogenous DNA sequence;
- a complementary strand oligonucleotide homologous to the target sequence;
- a nuclease capable of producing double stranded DNA blunt end cuts;
- wherein the exogenous DNA sequence comprises at least one nucleotide difference compared to the genome, both the target sequence and the target nucleic acid sequence in the genome are recognized and cut by the nuclease, and the recombinant vectors are configured such that that the target nucleic acid sequence in the genome is no longer present once the exogenous DNA sequence has been integrated into the genome in the correct orientation.
Dependencies are inferred from the claim text.