Triggered covalent probes for imaging and silencing genetic expression
The present invention relates to the use of cross-linking probes to covalently bind probes to nucleic acid targets. In some embodiments, the probe comprises an initiator region that is able to bind to a first portion of a target nucleic acid, a probe region linked too the initiator region that is able to bind to a second region of the target nucleic acid and that comprises one or more cross-linkers, and a blocking region hybridized to the probe region.
1. A cross-linking probe comprising:
a nucleic acid initiator region that is able to bind to a first portion of a target nucleic acid;
a nucleic acid probe region, wherein the probe region is linked to the initiator region and is able to bind to a second portion of the target nucleic acid that is contiguous with the first portion;
one or more cross-linkers that are part of the probe region, wherein the cross-linker is capable of forming a covalent bond to the target nucleic acid when the nucleic acid initiator region is bound to the first portion of the target nucleic acid and the probe region is bound to the second portion of the target nucleic acid; and
a blocking region that is hybridized to the probe region.
2. The cross-linking probe of claim 1 , wherein the initiator region is complementary to the first portion of the target nucleic acid.
3. The cross-linking probe of claim 1 , wherein the probe region is complementary to the second portion of the target nucleic acid.
4. The cross-linking probe of claim 1 , further comprising a loop region that links the probe region to the blocking region.
5. The cross-linking probe of claim 1 , wherein at least one cross-linker is an activatable cross-linker.
6. The cross-linking probe of claim 5 , wherein the activatable cross-linker is light activatable.
7. The cross-linking probe of claim 5 , wherein the activatable cross-linker is conformationally activatable.
8. The cross-linking probe of claim 1 , wherein at least one of the cross-linkers is selected from the group consisting of 4-Thio-dT and CNV-K.
9. The cross-linking probe of claim 1 , wherein the probe region comprises two or more cross linkers.
10. The cross-linking probe of claim 1 , wherein the probe region comprises at least one nucleotide on each side of the cross-linker.
11. The cross-linking probe of claim 1 , wherein the probe region is immediately adjacent to the initiator region.
12. The cross-linking probe of claim 1 , wherein the initiator region is a sticky end.