Methods of modifying a target nucleic acid with an argonaute
This disclosure provides for compositions and methods for the use of designed nucleic acid-targeting nucleic acids, Argonautes, and complexes thereof.
1. A method of cutting a double-stranded DNA target sequence in a host cell in vitro, wherein the double-stranded DNA target sequence comprises a first DNA strand and a second DNA strand, the method comprising: introducing into the host cell:
a first complex comprising a first TtArgonaute protein and a first guide nucleic acid, wherein the first guide nucleic acid hybridizes with the first strand of the double-stranded DNA target sequence in the host cell and the first complex cuts the first strand of the double-stranded DNA target sequence, and
a second complex comprising a second TtArgonaute protein and a second guide nucleic acid, wherein the second guide nucleic acid hybridizes with the second strand of the double-stranded DNA target sequence in the host cell and the second complex cuts the second strand of the double-stranded DNA target sequence.
2. The method of claim 1 , wherein the first strand cut and the second strand cut of the double-stranded DNA target sequence are such that the cutting of the double-stranded DNA target sequence is a blunt-end cut.
3. The method of claim 1 , wherein the first strand cut and the second strand cut of the double-stranded DNA taret sequence are such that the cutting of the double-stranded DNA target sequence is a staggared-end cut.
4. The method of claim 1 , wherein the first guide nucleic acid and the second guide nucleic acid comprise DNA.
5. The method of claim 1 , wherein the first guide nucleic acid and the second guide nucleic acid comprise RNA.
6. The method of claim 5 , wherein the first guide nucleic acid and the second guide nucleic acid further comprise DNA.
7. The method of claim 1 , wherein the double-stranded DNA target sequence is within a genomic DNA of a cukaryotic host cell.
8. The method of claim 1 , wherein the host cell is a eukaryotic cell.
9. The method of claim 1 , wherein the host cell is a plant cell, an algal cell, a fungal cell, a cell from a vertebrate animal, or a cell from a mammal.
10. The method of claim 1 , wherein the introducing is carried out ex vivo.
11. The method of claim 1 , wherein the double-stranded DNA target sequence comprises DNA, the first guide nucleic acid comprises DNA, and the second guid nucleic acid comprises DNA.
12. The method of claim 11 , wherein the host cell is a eukaryotic cell.