Light-sensitive ion-passing molecules

• US 9,604,073 B2
• Filed: 05/12/2016
• Issued: 03/28/2017
• Est. Priority Date: 03/17/2010
• Status: Active Grant

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Deisseroth, Karl, Zhang, Feng, Gradinaru, Viviana
• Primary Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US15/153,299
• Publication Number:
  US 20160287895A1
• Time in Patent Office: 320 Days
• Field of Search: 435/320.1, 530/350, 536/23.4
• US Class Current: 1/1
• CPC Class Codes: A01K 2207/05 Animals modified by non-integrating nucleic acids, e.g. antisense, RNAi, morpholino, episomal vector, for non-therapeutic purpose
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/0393 Animal model comprising a reporter system for screening tests
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61N 2005/0626 Monitoring, verifying, controlling systems and methods
• CPC Class Codes: A61N 2005/063 comprising light transmitting means, e.g. optical fibres
• CPC Class Codes: A61N 2005/0651 Diodes
• CPC Class Codes: A61N 2005/0663 Coloured light
• CPC Class Codes: A61N 5/0601 Apparatus for use inside the body
• CPC Class Codes: A61N 5/062 Photodynamic therapy, i.e. excitation of an agent
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 14/405 from algae
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: C07K 2319/01 containing a localisation/targetting motif
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 : having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C12N 15/86 : Viral vectors
• CPC Class Codes: C12N 2750/14143 : viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2799/025 : where the vector is derived from a parvovirus

Abstract

The invention provides polynucleotides and methods for expressing light-activated proteins in animal cells and altering an action potential of the cells by optical stimulation. The invention also provides animal cells and non-human animals comprising cells expressing the light-activated proteins.

Claims

1. A system comprising:

a) a delivery device comprising a polynucleotide that comprises a nucleotide sequence encoding a light-activated polypeptide, wherein the light-activated polypeptide comprises, from N-terminus to C-terminus;

i) a core amino acid sequence that is at least 95% identical to the sequence shown in SEQ ID NO;

3, SEQ ID NO;

1, SEQ ID NO;

2, or SEQ ID NO;

4;

ii) an endoplasmic reticulum (ER) export signal; and

iii) a membrane trafficking signal;

b) a light source; and

c) a control device that controls generation of light by the light source.

2. The system of claim 1, wherein the control device comprises a control circuit that is arranged to respond to an external signal.

3. The system of claim 1, wherein the control device comprises a control circuit that comprises one or more of a rectifier circuit, a battery, a pulse timer, and a comparator circuit.

4. The system of claim 1, wherein the control device comprises a control circuit that comprises an integrated circuit.

5. The system of claim 1, wherein all or a part of the system is implantable.

6. The system of claim 1, wherein the polynucleotide is present in a matrix.

7. The system of claim 1, wherein the polynucleotide is present in a gel.

8. The system of claim 1, wherein the light source comprises a light-emitting diode.

9. The system of claim 1, wherein the light source comprises a fiber optic.

10. The system of claim 1, wherein the ER export signal comprises the amino acid sequence FCEYENEV (SEQ ID NO:

12).

11. The system of claim 1, wherein the membrane trafficking signal comprises the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:

11).

12. The system of claim 1, wherein the core amino acid sequence is at least 95% identical to amino acids 25-265 of SEQ ID NO:

2.

13. The system of claim 1, wherein the core amino acid sequence is at least 95% identical to SEQ ID NO:

3.

14. The system of claim 1, wherein the core amino acid sequence is at least 95% identical to SEQ ID NO:

4.

15. The system of claim 1, wherein the light-activated protein-encoding nucleotide sequence is operably linked to a promoter.

16. The system of claim 15, wherein the promoter is a CaMKIIa promoter.

17. The system of claim 15, wherein the promoter is a synapsin I promoter.

18. The system of claim 1, wherein the polynucleotide is in an expression vector.

19. The system of claim 18, wherein the expression vector is a viral vector.

20. The system of claim 19, wherein the viral vector is an adenoassociated virus vector, a retroviral vector, an adenoviral vector, a herpes simplex virus vector, or a lentiviral vector.