Targeted transposition for use in epigenetic studies
Disclosed herein are compositions, methods and kits useful for epigenetic analysis based on the use of transposons that are targeted to specific regions of chromatin based on DNA-DNA interactions, protein-protein interactions, RNA-RNA interactions, and nucleic acid-protein interactions.
1. A binding protein-transposome complex, comprising a binding protein that binds a target, wherein the binding protein comprises Protein A, Protein G, a Protein A fusion protein or a Protein G fusion protein, said binding protein stably attached to a stable transposase:transposon complex (“transposome”), wherein the target is a protein that binds a nucleic acid-associated protein or protein domain, a DNA-binding site, or a protein that binds either of these.
2. The complex of claim 1 , further comprising one or more oligonucleotide bar code sequences to uniquely identify the binding protein.
3. The complex of claim 1 , further comprising an extraction moiety.
4. The complex of claim 3 , where the extraction moiety comprises a sequencing adaptor or PCR primer(s).
5. A binding protein-transposome complex, comprising a non- antibody protein, wherein the non-antibody protein comprises Protein A, Protein G, a Protein A fusion protein or a Protein G fusion protein, said protein stably attached to a stable transposase:transposon complex (“transposome”), wherein the non-antibody protein is further bound to a second protein, wherein the second protein binds a nucleic acid-associated protein or protein domain, or a DNA-binding site.
6. The complex of claim 5 , wherein the second protein comprises an antibody.
7. The composition of claim 1 , wherein the binding protein-transposome complex is bound, indirectly, to chromatin.
8. The composition of claim 1 , wherein the binding protein binds a molecule that binds to a nucleic acid-associated protein or protein domain, and wherein the molecule comprises an antibody, biotin or streptavidin.
9. A composition comprising:
(a) a primary antibody, wherein the primary antibody binds a nucleic acid-associated protein or protein domain, or a DNA-binding site;
(b) a secondary antibody bound to the primary antibody; and
(c) a binding protein-transposome complex, comprising a non-antibody protein bound to the secondary antibody, wherein the non-antibody protein comprises Protein A, Protein G, a Protein A fusion protein or a Protein G fusion protein, said non-antibody protein stably attached to a stable transposase:transposon complex (“transposome”).
10. The composition of claim 9 , wherein the primary antibody is directly bound to chromatin.
11. The complex of claim 1 , wherein the transposome is biotinylated, the binding protein is biotinylated, and the transposon and binding protein are conjugated through streptavidin.
12. The complex of claim 1 , wherein the transposome is biotinylated, and the binding protein bound to the transposon through a streptavidin-Protein A fusion protein or a streptavidin-Protein G fusion protein.
13. The complex of claim 1 , wherein the binding protein is conjugated to the transposome directly via a chemical crosslinker.
14. The complex of claim 5 , wherein the transposome is biotinylated, the non-antibody protein is biotinylated, and the transposon and binding protein are conjugated through streptavidin.
15. The complex of claim 5 , wherein the transposome is biotinylated, and the non-antibody protein comprises a streptavidin-Protein A fusion protein or a streptavidin-Protein G fusion protein.
16. The complex of claim 5 , wherein the binding protein is conjugated to the transposome directly via a chemical crosslinker.
17. The composition of claim 9 , wherein the transposome is biotinylated, the non-antibody protein is biotinylated, and the transposon and binding protein are conjugated through streptavidin.
18. The composition of claim 9 , wherein the transposome is biotinylated, and the non-antibody protein comprises a streptavidin-Protein A fusion protein or a streptavidin-Protein G fusion protein.
19. The composition of claim 9 , wherein the binding protein is conjugated to the transposome directly via a chemical crosslinker.
20. The complex of claim 5 , wherein the transposon further comprises one or more oligonucleotide bar code sequences to uniquely identify the second protein.
21. The complex of claim 9 , wherein the transposon further comprises one or more oligonucleotide bar code sequences to uniquely identify the primary antibody.
22. The complex of claim 5 , wherein the composition further comprises an extraction moiety.
23. The complex of claim 9 , wherein the composition further comprises an extraction moiety.
24. The complex of claim 6 , wherein the antibody is bound to chromatin.
25. The complex of claim 1 , wherein the target comprises an antibody that binds a nucleic acid-associated protein or protein domain, or a DNA-binding site.
26. The complex of claim 1 , wherein the target comprises a second antibody that binds a first antibody that binds a nucleic acid-associated protein or protein domain, or a DNA-binding site.