Methods of Light Activated Release of Ligands from Endosomes
Methods for delivering ligands to a cell by using light to activate fluorescent ligands causing their release from endosomes. The instant methods thus increasing the efficiency of ligands, e.g. in vitro or at localized sites within a subject. The invention provides for the release of ligands by shining a light source on a cell to promote release of ligands into the cell where they can effect their function.
1 . A method of delivering a ligand to a cell comprising:
a) contacting a cell with a ligand and a fluorophore; and
b) illuminating the cell with a light that activates the fluorophore such that the ligand is delivered to the cell.
2 . The method of claim 1 , wherein the ligand is an oligonucleotide.
3 . The method of claim 1 , wherein the ligand is peptide.
4 . The method of claim 1 , wherein ligand is a fluorescent virus.
5 . The method of claim 1 , wherein the ligand is a morpholino oligonucleotide.
6 . The method of claim 1 , wherein the ligand is a sense oligonucleotide.
7 . The method of claim 1 , wherein the ligand is an antisense oligonucleotide.
8 . The method of claim 1 , wherein the ligand enters an endosome of the cell during step (a).
9 . The method of claim 8 , wherein the illuminating of step (b) causes the endosome containing the ligand to release the ligand.
10 . The method of claim 1 , wherein the light has a wavelength of about 10 to about 380 nm.
11 . The method of claim 1 , wherein the light has a wavelength of about 380 to about 500 nm.
12 . The method of claim 1 , wherein the cells are illuminated for less than about 2 minutes.
13 . The method of claim 1 , wherein the cells are illuminated for less than about 1 minute.
14 . The method of claim 1 , wherein the light of step (b) is produced from a flexible endoscopic light source.
15 . The method of claim 1 , wherein the fluorophore and the ligand are linked via a covalent linkage.
16 . The method of claim 1 , wherein the fluorophore is a fluorescein fluorophore.
17 . The method of claim 1 , wherein the fluorophore and the ligand contacted with the cell simultaneously.
18 - 32 . (canceled)
33 . The method of claim 17 , wherein the localized site is inside the colon of the subject.
34 . The method of claim 17 , wherein the localized site is on the skin of the subject.
35 . The method of claim 17 , wherein the localized site is a tumor.
36 . A method of modulating protein production at a localized site in a subject comprising:
a) exposing a group of cells of the subject to a ligand and a fluorophore; and
b) illuminating the cells at a localized site in the subject with light that activates the fluorophore such that protein production at a localized site in the subject is modulated.
37 . The method of claim 36 , wherein protein production is enhanced.
38 . The method of claim 37 , wherein protein production is inhibited.
39 . A method of modulating protein activity at a localized site in a subject comprising:
a) exposing a group of cells of the subject to a ligand and a fluorophore; and
b) illuminating the cells at a localized site in the subject with light such that the fluorophore is activated and protein activity at a localized site in a subject is modulated.
40 . The method of claim 39 , wherein protein activity is enhanced.
41 . The method of claim 39 , wherein protein activity is inhibited.
42 . (canceled)