Optogenetic induction of neurodegenerative disease pathologies
View Patent ↗The present disclosure relates to compounds, compositions, and methods for the inducing neurodegenerative disease pathologies. In one aspect, disclosed herein is a nucleotide sequence encoding a chimeric polypeptide, comprising: a first nucleotide sequence encoding a light-induced oligomerization domain and a second nucleotide sequence encoding a neurodegenerative disease in target protein. Disclosed herein is a method of inducing a neurodegenerative disease pathology in a cell, comprising the steps: introducing into the cell an expression vector encoding a chimeric polypeptide, comprising: a first nucleotide sequence encoding a light-induced oligomerization domain and a second nucleotide sequence encoding a low complexity domain from a neurodegenerative disease target protein, wherein the first nucleotide sequence is operably linked to a promoter; expressing the chimeric polypeptide; and inducing oligomerization of the chimeric polypeptide by stimulation with blue light.
1. A nucleotide sequence encoding a chimeric polypeptide, comprising: a first nucleotide sequence encoding a light-induced oligomerization domain and a second nucleotide sequence encoding a low complexity domain from a neurodegenerative disease target protein; wherein the light-induced oligomerization domain is CRY2OLIG, or a fragment thereof and wherein the low complexity domain from a neurodegenerative disease target protein is TDP-43.
2. An expression vector encoding a chimeric polypeptide, the expression vector comprising: the nucleotide sequence of claim 1 .
3. An isolated cell comprising the nucleotide sequence of claim 1 .
4. A chimeric polypeptide comprising a light-induced oligomerization domain and a low complexity domain from a neurodegenerative disease target protein; wherein the light-induced oligomerization domain is CRY2OLIG, or a fragment thereof; and wherein the low complexity domain from a neurodegenerative disease target protein is TDP-43.
5. A method of inducing a neurodegenerative disease pathology in a cell, comprising the steps of:
introducing into the cell an expression vector encoding a chimeric polypeptide, the expression vector comprising: the nucleotide sequence of claim 1 , wherein the nucleotide sequence is operably linked to a promoter;
expressing the chimeric polypeptide; and
inducing oligomerization of the chimeric polypeptide by stimulation with blue light.
6. A method of screening for an agent that modulates protein aggregation, comprising the steps of:
introducing into a cell an expression vector encoding a chimeric polypeptide, the expression vector comprising: the nucleotide sequence of claim 1 , wherein the nucleotide sequence is operably linked to a promoter;
expressing the chimeric polypeptide;
introducing the agent into a culture media comprising the cell;
inducing oligomerization of the chimeric polypeptide by stimulation with blue light; and
determining modulation of protein aggregation by the agent.
7. The nucleotide sequence of claim 1 , wherein the first nucleotide sequence encodes a light-induced oligomerization domain comprising at least 90% sequence identity to SEQ ID NO: 2.
8. The nucleotide sequence of claim 1 , wherein the first nucleotide sequence encodes a light-induced oligomerization domain comprising SEQ ID NO: 2.
9. The nucleotide sequence of claim 1 , wherein the first nucleotide sequence and the second nucleotide sequence encode a chimeric polypeptide comprising at least 90% sequence identity to SEQ ID NO: 6.
10. The nucleotide sequence of claim 1 , wherein the first nucleotide sequence and the second nucleotide sequence encode a chimeric polypeptide comprising SEQ ID NO: 6.
11. The nucleotide sequence of claim 1 , wherein the first nucleotide sequence and the second nucleotide sequence encode a chimeric polypeptide comprising at least 90% sequence identity to SEQ ID NO: 7.
12. The nucleotide sequence of claim 1 , wherein the first nucleotide sequence and the second nucleotide sequence encode a chimeric polypeptide comprising SEQ ID NO: 7.
13. The nucleotide sequence of claim 1 , wherein the second nucleotide sequence encodes a low complexity domain comprising at least 90% sequence identity to SEQ ID NO: 95.
14. The nucleotide sequence of claim 1 , wherein the second nucleotide sequence encodes a low complexity domain comprising SEQ ID NO: 95.