Methods, compositions and kits for assaying mitochondrial function
US-2015198585-A1 · Jul 16, 2015 · US
Methods, compositions and kits for assaying mitochondrial function
US9915647B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9915647-B2 |
| Application number | US-201615087521-A |
| Country | US |
| Kind code | B2 |
| Filing date | Mar 31, 2016 |
| Priority date | Apr 8, 2011 |
| Publication date | Mar 13, 2018 |
| Grant date | Mar 13, 2018 |
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- Title
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Abstract
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The invention provides methods, compositions, devices, and kits relating to the use of cholesterol-dependent cytolysins (e.g., PFOs) for measuring intracellular mitochondrial activity.
First claim
Opening claim text (preview).
What is claimed is: 1. A method for introducing an exogenous agent into a permeabilized cell comprising: (a) incubating a cell with a cholesterol-dependent cytolysin that lacks cysteine residues and that lacks an N-terminal signal sequence, thereby selectively permeabilizing the plasma membrane of the cell; and (b) introducing an exogenous agent into the permeabilized cell. 2. The method of claim 1 , wherein the exogenous agent is a nucleic acid. 3. The method of claim 1 , wherein the exogenous agent is protein. 4. The method of claim 1 , wherein the exogenous agent is an antibody. 5. The method of claim 1 , wherein the exogenous agent is a dye or fluorescent protein. 6. The method of claim 1 , wherein the dye is a calcium sensitive dye or a pH-sensing dye. 7. The method of claim 1 , wherein the exogenous agent is a reducing agent, a respiratory chain inhibitor, an oxidative phosphorylation inhibitor, an uncoupling agent, a transport inhibitor, an ionophore or a Krebs cycle inhibitor. 8. The method of claim 1 , wherein the exogenous agent is a test agent. 9. The method of claim 1 , further comprising measuring the effect of the exogenous agent on the cell. 10. The method of claim 9 , wherein said measuring comprises measuring an intracellular function of the cell. 11. The method of claim 10 , wherein the intracellular function is a mitrochondrial function. 12. The method of claim 1 , wherein the cell is a mammalian cell. 13. The method of claim 1 , wherein the exogenous agent cannot cross the plasma membrane of a non-permeabilized cell. 14. The method of claim 1 , wherein the cholesterol-dependent cytolysin is a variant of a Perfringolysin O (PFO). 15. The method of claim 1 , wherein the exogenous agent is a substrate for a mitochondrial enzyme.
Assignees
Inventors
Classifications
from Clostridium (G) · CPC title
involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title
- G01N33/5079Primary
Mitochondria · CPC title
- C07K14/33Primary
from Clostridium (G) · CPC title
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Frequently asked questions
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- What does patent US9915647B2 cover?
- The invention provides methods, compositions, devices, and kits relating to the use of cholesterol-dependent cytolysins (e.g., PFOs) for measuring intracellular mitochondrial activity.
- Who is the assignee on this patent?
- Baystate Medical Center Inc, Univ Massachusetts
- What technology area does this patent fall under?
- Primary CPC classification G01N33/5079. Mapped technology areas include Physics.
- When was this patent published?
- Publication date Tue Mar 13 2018 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 1 related publication on this page (citations in our corpus or others sharing the same primary CPC).