JP2019522469A - チャネルロドプシンの変異型光誘導性イオンチャネル - Google Patents
チャネルロドプシンの変異型光誘導性イオンチャネル Download PDFInfo
- Publication number
- JP2019522469A JP2019522469A JP2018563484A JP2018563484A JP2019522469A JP 2019522469 A JP2019522469 A JP 2019522469A JP 2018563484 A JP2018563484 A JP 2018563484A JP 2018563484 A JP2018563484 A JP 2018563484A JP 2019522469 A JP2019522469 A JP 2019522469A
- Authority
- JP
- Japan
- Prior art keywords
- seq
- xaa
- ion channel
- light
- mutant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000004310 Ion Channels Human genes 0.000 title claims abstract description 105
- 108090000862 Ion Channels Proteins 0.000 title description 82
- 108010035848 Channelrhodopsins Proteins 0.000 title description 22
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 19
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 18
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 18
- 239000013604 expression vector Substances 0.000 claims abstract description 13
- 210000004027 cell Anatomy 0.000 claims description 119
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 96
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 95
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 75
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 52
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 50
- 108091006146 Channels Proteins 0.000 claims description 41
- 238000006467 substitution reaction Methods 0.000 claims description 33
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 31
- 235000001014 amino acid Nutrition 0.000 claims description 26
- 239000011780 sodium chloride Substances 0.000 claims description 26
- 239000000243 solution Substances 0.000 claims description 26
- 210000002569 neuron Anatomy 0.000 claims description 22
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims description 20
- 239000007995 HEPES buffer Substances 0.000 claims description 20
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 19
- 238000005259 measurement Methods 0.000 claims description 14
- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 claims description 13
- 238000013537 high throughput screening Methods 0.000 claims description 13
- 230000000638 stimulation Effects 0.000 claims description 12
- 241001465754 Metazoa Species 0.000 claims description 9
- 208000003098 Ganglion Cysts Diseases 0.000 claims description 7
- 208000005400 Synovial Cyst Diseases 0.000 claims description 7
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims description 6
- 210000004962 mammalian cell Anatomy 0.000 claims description 5
- 239000004475 Arginine Substances 0.000 claims description 4
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 4
- 235000004279 alanine Nutrition 0.000 claims description 4
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 4
- 210000004978 chinese hamster ovary cell Anatomy 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 210000000608 photoreceptor cell Anatomy 0.000 claims description 4
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims description 3
- 239000004472 Lysine Substances 0.000 claims description 3
- 206010029260 Neuroblastoma Diseases 0.000 claims description 3
- 125000000539 amino acid group Chemical group 0.000 claims description 3
- 210000004565 granule cell Anatomy 0.000 claims description 3
- 230000000971 hippocampal effect Effects 0.000 claims description 3
- 210000000449 purkinje cell Anatomy 0.000 claims description 3
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 claims description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 2
- 235000003704 aspartic acid Nutrition 0.000 claims description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 2
- 210000001052 bipolar neuron Anatomy 0.000 claims description 2
- 235000013922 glutamic acid Nutrition 0.000 claims description 2
- 239000004220 glutamic acid Substances 0.000 claims description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 2
- 201000000050 myeloid neoplasm Diseases 0.000 claims description 2
- 210000001510 pseudounipolar neuron Anatomy 0.000 claims description 2
- 239000002773 nucleotide Substances 0.000 claims 2
- 125000003729 nucleotide group Chemical group 0.000 claims 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims 1
- 239000004473 Threonine Substances 0.000 claims 1
- 239000002253 acid Substances 0.000 claims 1
- 210000003078 multipolar neuron Anatomy 0.000 claims 1
- 230000001976 improved effect Effects 0.000 abstract description 3
- 150000001413 amino acids Chemical class 0.000 description 21
- 239000011575 calcium Substances 0.000 description 20
- 210000003050 axon Anatomy 0.000 description 16
- 239000012528 membrane Substances 0.000 description 14
- 238000000034 method Methods 0.000 description 13
- 230000002207 retinal effect Effects 0.000 description 12
- NCYCYZXNIZJOKI-UHFFFAOYSA-N vitamin A aldehyde Natural products O=CC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-UHFFFAOYSA-N 0.000 description 12
- 230000010221 calcium permeability Effects 0.000 description 11
- 239000000126 substance Substances 0.000 description 11
- 210000001787 dendrite Anatomy 0.000 description 10
- 210000001525 retina Anatomy 0.000 description 9
- 108020004414 DNA Proteins 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 230000014509 gene expression Effects 0.000 description 7
- 230000035772 mutation Effects 0.000 description 7
- 230000004044 response Effects 0.000 description 7
- 230000000007 visual effect Effects 0.000 description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- 230000004913 activation Effects 0.000 description 6
- 210000004899 c-terminal region Anatomy 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 6
- 230000001537 neural effect Effects 0.000 description 6
- 210000001743 on-bipolar cell Anatomy 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 108091028043 Nucleic acid sequence Proteins 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 210000004556 brain Anatomy 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000001415 gene therapy Methods 0.000 description 5
- 230000001965 increasing effect Effects 0.000 description 5
- 230000035699 permeability Effects 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 210000000225 synapse Anatomy 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- 108091026890 Coding region Proteins 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- 206010034972 Photosensitivity reaction Diseases 0.000 description 4
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 4
- 239000007983 Tris buffer Substances 0.000 description 4
- 210000000411 amacrine cell Anatomy 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 210000003984 auditory pathway Anatomy 0.000 description 4
- 210000000170 cell membrane Anatomy 0.000 description 4
- 239000011035 citrine Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 4
- 108091008695 photoreceptors Proteins 0.000 description 4
- 230000036211 photosensitivity Effects 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 210000003994 retinal ganglion cell Anatomy 0.000 description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 4
- 241000702421 Dependoparvovirus Species 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- 108010052285 Membrane Proteins Proteins 0.000 description 3
- 102000004257 Potassium Channel Human genes 0.000 description 3
- 241000283984 Rodentia Species 0.000 description 3
- 241000195615 Volvox Species 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- 210000001153 interneuron Anatomy 0.000 description 3
- 210000003292 kidney cell Anatomy 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- 108020001213 potassium channel Proteins 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 150000003726 retinal derivatives Chemical class 0.000 description 3
- 108091005957 yellow fluorescent proteins Proteins 0.000 description 3
- NCYCYZXNIZJOKI-HPNHMNAASA-N 11Z-retinal Natural products CC(=C/C=O)C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-HPNHMNAASA-N 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- 241000699800 Cricetinae Species 0.000 description 2
- 241000252212 Danio rerio Species 0.000 description 2
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- 208000032041 Hearing impaired Diseases 0.000 description 2
- 108010092555 Large-Conductance Calcium-Activated Potassium Channels Proteins 0.000 description 2
- 102000016469 Large-Conductance Calcium-Activated Potassium Channels Human genes 0.000 description 2
- 241000282560 Macaca mulatta Species 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 241000276569 Oryzias latipes Species 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 241000555745 Sciuridae Species 0.000 description 2
- 241000611306 Taeniopygia guttata Species 0.000 description 2
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 2
- 108700005077 Viral Genes Proteins 0.000 description 2
- 229930002945 all-trans-retinaldehyde Natural products 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 229910001424 calcium ion Inorganic materials 0.000 description 2
- 210000005056 cell body Anatomy 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 2
- 238000010304 firing Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- YFHXZQPUBCBNIP-UHFFFAOYSA-N fura-2 Chemical compound CC1=CC=C(N(CC(O)=O)CC(O)=O)C(OCCOC=2C(=CC=3OC(=CC=3C=2)C=2OC(=CN=2)C(O)=O)N(CC(O)=O)CC(O)=O)=C1 YFHXZQPUBCBNIP-UHFFFAOYSA-N 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 238000001476 gene delivery Methods 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 210000004295 hippocampal neuron Anatomy 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 230000028161 membrane depolarization Effects 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000007420 reactivation Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000001116 retinal neuron Anatomy 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910001415 sodium ion Inorganic materials 0.000 description 2
- 210000001082 somatic cell Anatomy 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 210000000278 spinal cord Anatomy 0.000 description 2
- 210000001323 spiral ganglion Anatomy 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000002834 transmittance Methods 0.000 description 2
- NCYCYZXNIZJOKI-IOUUIBBYSA-N 11-cis-retinal Chemical compound O=C/C=C(\C)/C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-IOUUIBBYSA-N 0.000 description 1
- NCYCYZXNIZJOKI-HWCYFHEPSA-N 13-cis-retinal Chemical compound O=C/C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-HWCYFHEPSA-N 0.000 description 1
- WWRGFLIMFQYXRS-UHFFFAOYSA-N 3,7,11-trimethyldodeca-2,4,6,8,10-pentaenal Chemical compound CC(C)=CC=CC(C)=CC=CC(C)=CC=O WWRGFLIMFQYXRS-UHFFFAOYSA-N 0.000 description 1
- MIMKOTXYWWOEFT-UHFFFAOYSA-N 3,7-dimethyldeca-2,4,6,8-tetraenal Chemical compound CC=CC(C)=CC=CC(C)=CC=O MIMKOTXYWWOEFT-UHFFFAOYSA-N 0.000 description 1
- UDYGWSPUYSRWRN-UHFFFAOYSA-N 3,7-dimethylocta-2,4,6-trienal Chemical compound CC(C)=CC=CC(C)=CC=O UDYGWSPUYSRWRN-UHFFFAOYSA-N 0.000 description 1
- QPRQNCDEPWLQRO-UHFFFAOYSA-N 3R-hydroxy-all-trans-retinal Natural products O=CC=C(C)C=CC=C(C)C=CC1=C(C)CC(O)CC1(C)C QPRQNCDEPWLQRO-UHFFFAOYSA-N 0.000 description 1
- URNCTMHGJQSLOC-UHFFFAOYSA-N 4-[4-[(1z,3e)-4-[4-(dipentylamino)phenyl]buta-1,3-dienyl]pyridin-1-ium-1-yl]butane-1-sulfonate Chemical compound C1=CC(N(CCCCC)CCCCC)=CC=C1C=CC=CC1=CC=[N+](CCCCS([O-])(=O)=O)C=C1 URNCTMHGJQSLOC-UHFFFAOYSA-N 0.000 description 1
- NCYCYZXNIZJOKI-MKOSUFFBSA-N 9-cis-retinal Chemical compound O=C/C=C(\C)/C=C/C=C(/C)\C=C\C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-MKOSUFFBSA-N 0.000 description 1
- 241000710929 Alphavirus Species 0.000 description 1
- 241000252073 Anguilliformes Species 0.000 description 1
- 241000258141 Arbacia punctulata Species 0.000 description 1
- 239000000592 Artificial Cell Substances 0.000 description 1
- 241000244203 Caenorhabditis elegans Species 0.000 description 1
- 241000282465 Canis Species 0.000 description 1
- 108091005462 Cation channels Proteins 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 241000700199 Cavia porcellus Species 0.000 description 1
- 241000282552 Chlorocebus aethiops Species 0.000 description 1
- 241000867607 Chlorocebus sabaeus Species 0.000 description 1
- 241000251571 Ciona intestinalis Species 0.000 description 1
- 108700010070 Codon Usage Proteins 0.000 description 1
- 108091035707 Consensus sequence Proteins 0.000 description 1
- 241001653748 Doryteuthis pealeii Species 0.000 description 1
- 241000255601 Drosophila melanogaster Species 0.000 description 1
- 241000257465 Echinoidea Species 0.000 description 1
- 241000283070 Equus zebra Species 0.000 description 1
- 241000238376 Euprymna scolopes Species 0.000 description 1
- 241000282324 Felis Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 206010019899 Hereditary retinal dystrophy Diseases 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101001032837 Homo sapiens Metabotropic glutamate receptor 6 Proteins 0.000 description 1
- 241000243251 Hydra Species 0.000 description 1
- 229930195714 L-glutamate Natural products 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 102100038300 Metabotropic glutamate receptor 6 Human genes 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 235000009421 Myristica fragrans Nutrition 0.000 description 1
- 108091061960 Naked DNA Proteins 0.000 description 1
- 241000282579 Pan Species 0.000 description 1
- 241000282520 Papio Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 206010034960 Photophobia Diseases 0.000 description 1
- 241000530496 Pristionchus pacificus Species 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 241000711798 Rabies lyssavirus Species 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- NCYCYZXNIZJOKI-OVSJKPMPSA-N Retinaldehyde Chemical compound O=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-OVSJKPMPSA-N 0.000 description 1
- 239000002262 Schiff base Substances 0.000 description 1
- 150000004753 Schiff bases Chemical class 0.000 description 1
- 241000710961 Semliki Forest virus Species 0.000 description 1
- 241000144290 Sigmodon hispidus Species 0.000 description 1
- 241000258128 Strongylocentrotus purpuratus Species 0.000 description 1
- 241001656718 Symsagittifera roscoffensis Species 0.000 description 1
- 241001441723 Takifugu Species 0.000 description 1
- 241001441724 Tetraodontidae Species 0.000 description 1
- 206010047571 Visual impairment Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- QPRQNCDEPWLQRO-DAWLFQHYSA-N all-trans-3-Hydroxyretinal Chemical compound O=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CC(O)CC1(C)C QPRQNCDEPWLQRO-DAWLFQHYSA-N 0.000 description 1
- CYVVUYORRQQAQE-RMWYGNQTSA-N all-trans-4-hydroxyretinal Chemical compound O=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)C(O)CCC1(C)C CYVVUYORRQQAQE-RMWYGNQTSA-N 0.000 description 1
- 125000000746 allylic group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 210000002226 anterior horn cell Anatomy 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000000823 artificial membrane Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 239000005441 aurora Substances 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 230000027288 circadian rhythm Effects 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 230000001054 cortical effect Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 238000013480 data collection Methods 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 239000000412 dendrimer Substances 0.000 description 1
- 229920000736 dendritic polymer Polymers 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- HUPFGZXOMWLGNK-UHFFFAOYSA-N diflunisal Chemical compound C1=C(O)C(C(=O)O)=CC(C=2C(=CC(F)=CC=2)F)=C1 HUPFGZXOMWLGNK-UHFFFAOYSA-N 0.000 description 1
- 229960000616 diflunisal Drugs 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000002964 excitative effect Effects 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- VPSRLGDRGCKUTK-UHFFFAOYSA-N fura-2-acetoxymethyl ester Chemical compound CC(=O)OCOC(=O)CN(CC(=O)OCOC(C)=O)C1=CC=C(C)C=C1OCCOC(C(=C1)N(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)=CC2=C1OC(C=1OC(=CN=1)C(=O)OCOC(C)=O)=C2 VPSRLGDRGCKUTK-UHFFFAOYSA-N 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000002267 hypothalamic effect Effects 0.000 description 1
- 210000003016 hypothalamus Anatomy 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004941 influx Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- QRXWMOHMRWLFEY-UHFFFAOYSA-N isoniazide Chemical compound NNC(=O)C1=CC=NC=C1 QRXWMOHMRWLFEY-UHFFFAOYSA-N 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000003127 knee Anatomy 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 208000013469 light sensitivity Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000001115 mace Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 241001515942 marmosets Species 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 238000012895 mono-exponential function Methods 0.000 description 1
- 210000002161 motor neuron Anatomy 0.000 description 1
- 230000004118 muscle contraction Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000000715 neuromuscular junction Anatomy 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000002831 pharmacologic agent Substances 0.000 description 1
- 230000002186 photoactivation Effects 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 150000004291 polyenes Chemical class 0.000 description 1
- 230000001242 postsynaptic effect Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 210000001176 projection neuron Anatomy 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000011241 protective layer Substances 0.000 description 1
- 210000001747 pupil Anatomy 0.000 description 1
- 230000001179 pupillary effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000002336 repolarization Effects 0.000 description 1
- 210000000964 retinal cone photoreceptor cell Anatomy 0.000 description 1
- 210000000880 retinal rod photoreceptor cell Anatomy 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 210000001908 sarcoplasmic reticulum Anatomy 0.000 description 1
- 210000001044 sensory neuron Anatomy 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000010473 stable expression Effects 0.000 description 1
- 230000000153 supplemental effect Effects 0.000 description 1
- 210000000221 suprachiasmatic nucleus Anatomy 0.000 description 1
- 210000001103 thalamus Anatomy 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 102000035160 transmembrane proteins Human genes 0.000 description 1
- 108091005703 transmembrane proteins Proteins 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 241001529453 unidentified herpesvirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 208000029257 vision disease Diseases 0.000 description 1
- 230000004393 visual impairment Effects 0.000 description 1
- 210000004127 vitreous body Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/405—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from algae
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/30—Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0618—Cells of the nervous system
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2510/00—Genetically modified cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Cell Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Virology (AREA)
- Microbiology (AREA)
- Ophthalmology & Optometry (AREA)
- Toxicology (AREA)
- Developmental Biology & Embryology (AREA)
- Neurology (AREA)
- Epidemiology (AREA)
- Neurosurgery (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Dermatology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
該変異型光誘導性イオンチャネルは、親光誘導性イオンチャネルと、配列番号1のF219に相当する位置における置換のみ異なっており、
前記置換は、-60 mVのクランプ電位、140 mM NaCl、2mM CaCl2、2MgCl2、10 mM HEPES、pH 7.4のバス溶液、及び110 mM NaCl、2 mM MgCl2、10 mM EGTA、10 mM HEPES、pH 7.4のピペット溶液で、全細胞構成における(in the whole cell configuration)パッチクランプ測定により比較したとき、前記親チャネルと比べて、前記変異型チャネルのオフキネティクスを促進する、変異型光誘導性イオンチャネルを開示する。
該変異型光誘導性イオンチャネルは、親光誘導性イオンチャネルと、配列番号2の位置F214に相当する位置における置換のみ異なっており、
前記置換は、-60 mVのクランプ電位、140 mM NaCl、2mM CaCl2、2MgCl2、10 mM HEPES、pH 7.4のバス溶液、及び110 mM NaCl、2 mM MgCl2、10 mM EGTA、10 mM HEPES、pH 7.4のピペット溶液で、全細胞構成におけるパッチクランプ測定により比較したとき、前記親チャネルと比べて、前記変異型チャネルのオフキネティクスを促進する、変異型光誘導性イオンチャネルを開示する。
該変異型光誘導性イオンチャネルは、親光誘導性イオンチャネルと、配列番号1の位置F259に相当する位置における置換のみ異なっており、
前記置換は、-60 mVのクランプ電位、140 mM NaCl、2mM CaCl2、2MgCl2、10 mM HEPES、pH 7.4のバス溶液、及び110 mM NaCl、2 mM MgCl2、10 mM EGTA、10 mM HEPES、pH 7.4のピペット溶液で、全細胞構成におけるパッチクランプ測定により比較したとき、前記親チャネルと比べて、前記変異型チャネルのオフキネティクスを促進する、変異型光誘導性イオンチャネルを開示する。
該変異型光誘導性イオンチャネルは、親光誘導性イオンチャネルと、配列番号1のF219に相当する位置における置換のみ異なっており、
前記置換は、特許請求の範囲にて規定されるような、-60 mVのクランプ電位、140 mM NaCl、2mM CaCl2、2MgCl2、10 mM HEPES、pH 7.4のバス溶液、及び110 mM NaCl、2 mM MgCl2、10 mM EGTA、10 mM HEPES、pH 7.4のピペット溶液で、全細胞構成におけるパッチクランプ測定により比較したとき、前記親チャネルと比べて、前記変異型チャネルのオフキネティクスを促進する。
該変異型光誘導性イオンチャネルは、親光誘導性イオンチャネルと、配列番号2の位置F214に相当する位置における置換のみ異なっており、
前記置換は、-60 mVのクランプ電位、140 mM NaCl、2mM CaCl2、2MgCl2、10 mM HEPES、pH 7.4のバス溶液、及び110 mM NaCl、2 mM MgCl2、10 mM EGTA、10 mM HEPES、pH 7.4のピペット溶液で、全細胞構成におけるパッチクランプ測定により比較したとき、前記親チャネルと比べて、前記変異型チャネルのオフキネティクスを促進する、変異型光誘導性イオンチャネルも記載する。好ましくは、前記置換は、F214Yである。
該変異型光誘導性イオンチャネルは、親光誘導性イオンチャネルと、配列番号3の位置F259に相当する位置における置換のみ異なっており、
前記置換は、-60 mVのクランプ電位、140 mM NaCl、2mM CaCl2、2MgCl2、10 mM HEPES、pH 7.4のバス溶液、及び110 mM NaCl、2 mM MgCl2、10 mM EGTA、10 mM HEPES、pH 7.4のピペット溶液で、全細胞構成におけるパッチクランプ測定により比較したとき、前記親チャネルと比べて、前記変異型チャネルのオフキネティクスを促進する、変異型光誘導性イオンチャネルも開示する。好ましくは、前記置換は、F259Yである。
Cys-Arg-Xaa3-Xaa4-Val-Xaa6-Xaa7-Met-Ala-Trp-Xaa11-Tyr-Phe-Val-Xaa15-Trp-Gly-Met-Phe-Pro-Xaa21-Leu-Phe-Xaa24-Leu
のモチーフ、
ここでXaa3はGln又はGluである、好ましくはここでXaa3はGlnである;
ここでXaa4はVal又はLeuである、好ましくはここでXaa4はValである;
ここでXaa6はThr又はArgである、好ましくはここでXaa6はThrである;
ここでXaa7はGly、Val又はAlaである、好ましくはここでXaa7はGlyである;
ここでXaa11はLeu又はThrである、好ましくはここでXaa11はLeuである;
ここでXaa15はSer又はAlaである、好ましくはここでXaa15はSerである;
ここでXaa21はIle又はValである、好ましくはここでXaa21はIleである;および
ここでXaa24はIle又はLeuである、好ましくはここでXaa24はIleである、
を含む。
Cys 132 + Asp 253; Cys 132 + Lys 257; Cys 132 + Trp 260; Cys 132 + Glu 123; Cys 132 + His 134; Cys 132 + Arg 134; Cys 132 + Thr 128; Cys 132 + Ser 128; Cys 132 + Ala 128; Cys 132 + Ala 156;
Cys 132 + Asp 253 + Lys 257; Cys 132 + Asp 253 + Trp 260; Cys 132 + Asp 253 + Glu 123; Cys 132 + Asp 253 + His 134; Cys 132 + Asp 253 + Arg 134; Cys 132 + Asp 253 + Thr 128; Cys 132 + Asp 253 + Ser 128; Cys 132 + Asp 253 + Ala 128; Cys 132 + Asp 253 + Ala 156;
Cys 132 + Lys 257 + Trp 260; Cys 132 + Lys 257 + Glu 123; Cys 132 + Lys 257 + His 134; Cys 132 + Lys 257 + Arg 134; Cys 132 + Lys 257 + Thr 128; Cys 132 + Lys 257 + Ser 128; Cys 132 + Lys 257 + Ala 128; Cys 132 + Lys 257 + Ala 156;
Cys 132 + Trp 260 + Glu 123; Cys 132 + Trp 260 + His 134; Cys 132 + Trp 260 + Arg 134; Cys 132 + Trp 260 + Thr 128; Cys 132 + Trp 260 + Ser 128; Cys 132 + Trp 260 + Ala 128; Cys 132 + Trp 260 + Ala 156;
Cys 132 + Glu 123 + His 134; Cys 132 + Glu 123 + His 134; Cys 132 + Glu 123 + Arg 134; Cys 132 + Glu 123 + Thr 128; Cys 132 + Glu 123 + Ser 128; Cys 132 + Glu 123 + Ala 128; Cys 132 + Glu 123 + Ala 156;
Cys 132 + His 134 + Thr 128; Cys 132 + His 134 + Ser 128; Cys 132 + His 134 + Ala 128; Cys 132 + His 134 + Ala 156;
Cys 132 + Arg 134 + Thr 128; Cys 132 + Arg 134 + Ser 128; Cys 132 + Arg 134 + Ala 128; Cys 132 + Arg 134 + Ala 156;
Cys 132 + Thr 128 + Ala 156; Cys 132 + Ser 128 + Ala 156; Cys 132 + Ala 128 + Ala 156;
Cys 132 + Asp 253 + Lys 257 + Trp 260; Cys 132 + Asp 253 + Lys 257 + Glu 123; Cys 132 + Asp 253 + Lys 257 + His 134; Cys 132 + Asp 253 + Lys 257 + Arg 134; Cys 132 + Asp 253 + Lys 257 + Thr 128; Cys 132 + Asp 253 + Lys 257 + Ser 128; Cys 132 + Asp 253 + Lys 257 + Ala 128; Cys 132 + Asp 253 + Lys 257 + Ala 156;
Cys 132 + Lys 157 + Trp 260 + Glu 123; Cys 132 + Lys 157 + Trp 260 + His 134; Cys 132 + Lys 157 + Trp 260 + Arg 134; Cys 132 + Lys 157 + Trp 260 + Thr 128; Cys 132 + Lys 157 + Trp 260 + Ser 128; Cys 132 + Lys 157 + Trp 260 + Ala 128; Cys 132 + Lys 157 + Trp 260 + Ala 156;
Cys 132 + Trp 260 + Glu 123 + His 134; Cys 132 + Trp 260 + Glu 123 +Arg 134; Cys 132 + Trp 260 + Glu 123 + Thr 128; Cys 132 + Trp 260 + Glu 123 + Ser 128; Cys 132 + Trp 260 + Glu 123 + Ala 128; Cys 132 + Trp 260 + Glu 123 + Ala 156;
Cys 132 + Glu 123 + His 134 + Thr 128; Cys 132 + Glu 123 + His 134 + Ser 128; Cys 132 + Glu 123 + His 134 + Ala 128; Cys 132 + Glu 123 + His 134 + Ala 156;
Cys 132 + Glu 123 + Arg 134 + Thr 128; Cys 132 + Glu 123 + Arg 134 + Ser 128; Cys 132 + Glu 123 + Arg 134 + Ala 128; Cys 132 + Glu 123 + Arg 134 + Ala 156;
Cys 132 + His 134 + Thr 128 + Ala 156; Cys 132 + His 134 + Ser 128 + Ala 156; Cys 132 + His 134 + Ala 128 + Ala 156;
Cys 132 + Arg 134 + Thr 128 + Ala 156; Cys 132 + Arg 134 + Ser 128 + Ala 156; Cys 132 + Arg 134 + Ala 128 + Ala 156;
Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123; Cys 132 + Asp 253 + Lys 257 + Trp 260 + His 134; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Arg 134; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Thr 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Ser 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Ala 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Ala 156;
Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Thr 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Ser 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Ala 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Ala 156;
Cys 132 + Trp 260 + Glu 123 + His 134 + Thr 128; Cys 132 + Trp 260 + Glu 123 + His 134 + Ser 128; Cys 132 + Trp 260 + Glu 123 + His 134 + Ala 128; Cys 132 + Trp 260 + Glu 123 + His 134 + Ala 156;
Cys 132 + Trp 260 + Glu 123 + Arg 134 + Thr 128; Cys 132 + Trp 260 + Glu 123 + Arg 134 + Ser 128; Cys 132 + Trp 260 + Glu 123 + Arg 134 + Ala 128; Cys 132 + Trp 260 + Glu 123 + Arg 134 + Ala 156;
Cys 132 + Glu 123 + Arg 134 + Thr 128 + Ala 156; Cys 132 + Glu 123 + Arg 134 + Ser 128 + Ala 156; Cys 132 + Glu 123 + Arg 134 + Ala 128 + Ala 156;
Cys 132 + Glu 123 + His 134 + Thr 128 + Ala 156; Cys 132 + Glu 123 + His 134 + Ser 128 + Ala 156; Cys 132 + Glu 123 + His 134 + Ala 128 + Ala 156;
Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Thr 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Ser 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Ala 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Ala 156;
Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134 + Thr 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ser 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ala 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ala 156;
Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Thr 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ser 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ala 128; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ala 156;
Cys 132 + Trp 260 + Glu 123 + Arg 134 + Thr 128 + Ala 156; Cys 132 + Trp 260 + Glu 123 + Arg 134 + Ser 128 + Ala 156; Cys 132 + Trp 260 + Glu 123 + Arg 134 + Ala 128 + Ala 156;
Cys 132 + Trp 260 + Glu 123 + His 134 + Thr 128 + Ala 156; Cys 132 + Trp 260 + Glu 123 + His 134 + Ser 128 + Ala 156; Cys 132 + Trp 260 + Glu 123 + His 134 + Ala 128 + Ala 156;
Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134 + Thr 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ser 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ala 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ala 156;
Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Thr 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ser 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ala 128; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ala 156;
Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134 + Thr 128 + Ala 156; Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ser 128 + Ala 156; Cys 132 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ala 128 + Ala 156;
Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Thr 128 + Ala 156; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ser 128 + Ala 156; Cys 132 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ala 128 + Ala 156;
Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134 + Thr 128 + Ala 156; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ser 128 + Ala 156; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + His 134 + Ala 128 + Ala 156;
Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Thr 128 + Ala 156; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ser 128 + Ala 156; Cys 132 + Asp 253 + Lys 257 + Trp 260 + Glu 123 + Arg 134 + Ala 128 + Ala 156。
Cys-Arg-Xaa3-Xaa4-Val-Xaa6-Xaa7-Met-Ala-Trp-Xaa11-Tyr-Phe-Val-Xaa15-Trp-Gly-Met-Phe-Pro-Xaa21-Leu-Phe-Xaa24-Leu、
ここでXaa3はGln又はGluである、好ましくはここでXaa3はGlnである;
ここでXaa4はVal又はLeuである、好ましくはここでXaa4はValである;
ここでXaa6はThr又はArgである、好ましくはここでXaa6はThrである;
ここでXaa7はGly、Val又はAlaである、好ましくはここでXaa7はGlyである;
ここでXaa11はLeu又はThrである、好ましくはここでXaa11はLeuである;
ここでXaa15はSer又はAlaである、好ましくはここでXaa15はSerである;
ここでXaa21はIle又はValである、好ましくはここでXaa21はIleである;かつ
ここでXaa24はIle又はLeuである、好ましくはここでXaa24はIleである。
Xaa1-Asp-Xaa3-Xaa4-Xaa5-Lys-Xaa7-Xaa8-Xaa9
ここでXaa1は、Leu、Ile、Ala、又はCysである;
ここでXaa3、Xaa4、Xaa5、Xaa7、及びXaa8 は、独立して任意のアミノ酸である;
ここでXaa9 は、Thr、Phe、又はTyrである。
本発明者らの目的は、変異によりオフキネティクスを更に加速することができるChR-2の第6膜貫通ドメイン内の残基を同定することであった。本発明者らは、第6膜貫通ドメインに焦点を当てた。
Hernandez et al. J Clin Invest. 2014;124(3):1114-29は、げっ歯類において聴覚経路の光遺伝学的刺激のための戦略を示した。特に、この著者らは、光ゲートされたイオンチャネルチャネルロドプシン-2(ChR2)を発現するように遺伝子操作された神経細胞の光刺激となる光遺伝学的刺激を特徴付けるための動物モデルについて説明している。螺旋神経節細胞(SGN)の光遺伝学的刺激は、単一の神経細胞及び神経細胞の集団応答の記録により実証されるように、聴覚経路を活性化する。さらに、SGNの光遺伝学的刺激は、聴覚障害マウスにおける聴覚活性を回復させる。閾値を超える光学的、音響的、および電気的刺激(光遺伝学的刺激を示す)に応答する下丘内の局所電場電位(local field potentials:LFP)を記録することによる蝸牛の興奮の空間的広がりを近似することにより、単極電気刺激よりも良好な周波数分解能が達成される。
Mace et al. Mol Ther. 2015;23(1):7-16は、アデノ随伴ウイルス(AAV)遺伝子治療による網膜神経細胞の光遺伝学的再活性化を記載する、本発明者らの一部によって執筆された初期の刊行物である。ほとんどの遺伝性網膜ジストロフィーでは、重度の視覚障害を引き起こす進行性の光受容体細胞変性を示す。アデノ随伴ウイルス(AAV)遺伝子治療による網膜神経細胞の光遺伝学的再活性化は、患者特異的な変異にかかわらず視力を回復させる可能性がある。臨床的な翻訳可能性への挑戦は、脆弱な変性網膜のために外科的に安全な送達経路を用いながらできるだけ自然な視力に近い視力を回復することである。網膜内側の視覚処理を維持するために、疾患の後期の段階でも依然として存在するON双極細胞が標的とされる。安全な遺伝子送達のために、眼に容易にアクセス可能な硝子体液への注入後に双極細胞を形質導入することができる近年設計されたAAV変異体が使用される。ON双極細胞特異的プロモーター下でチャネルロドプシンをコードするAAVは、硝子体内投与後にON双極細胞に限定的な長期の遺伝子送達を仲介することが示されている。ON双極細胞におけるチャネルロドプシン発現は、網膜レベルおよび皮質レベルでのONおよびOFF応答の回復をもたらす。さらに、光誘発性の運動器官の挙動が、治療を受けた盲目マウスにおいて回復される。
US 8,759,492 B2
WO 03/084994
WO 2012/032103
WO 2013/071231
1. Nagel, G. et al. Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc Natl Acad Sci USA 100, 13940-13945 (2003).
2. Nagel, G. et al. Light activation of channelrhodopsin-2 in excitable cells of Caenorhabditis elegans triggers rapid behavioral responses. Curr Biol 15, 2279-2284 (2005).
3. Boyden, E., Zhang, F., Bamberg, E., Nagel, G. & Deisseroth, K. Millisecond-timescale, genetically targeted optical control of neural activity. Nat Neurosci 8, 1263-1268 (2005).
4. Zhang, F. et al. Multimodal fast optical interrogation of neural circuitry. Nature 446, 633-639 (2007).
5. Nagel, G. et al. Channelrhodopsin-1: A Light-Gated Proton Channel in Green Algae. Science. 296, 2395-2398 (2002).
6. Bamann, C., Gueta, R., Kleinlogel, S., Nagel, G. & Bamberg, E. Structural guidance of the photocycle of channelrhodopsin-2 by an interhelical hydrogen bond. Biochemistry 49, 267-278 (2010).
7. Berndt, A., Yizhar, O., Gunaydin, L., Hegemann, P. & Deisseroth, K. Bi-stable neural state switches. Nat Neurosci 12, 229-234 (2009).
8. Lin, J., Lin, M., Steinbach, P. & Tsien, R. Characterization of engineered channelrhodopsin variants with improved properties and kinetics. Biophys J 96, 1803-1814 (2009).
9. Klapoetke N. et al. Independent optical excitation of distinct neural populations. Nature Methods. 11, 338-346 (2014).
10. Zhang F. et al. Red-shifted optogenetic excitation: a tool for fast neural control derived from Volvox carteri. Nature Neuroscience. 11, 631-633 (2008).
11. Lin J. Y. et al. ReaChR: a red-shifted variant of channelrhodopsin enables deep transcranial optogenetic excitation. Nature Neuroscience. 16, 1499-1508 (2013).
12. Kleinlogel S. et al. Ultra light-sensitive and fast neuronal activation with the Ca-permeable channelrhodopsin CatCh. Nature Neuroscience. 14, 513-518 (2011).
13. Hernandez et al. Optogenetic stimulation of the auditory pathway. J Clin Invest. 124(3): 1114-1129 (2014).
14. Mace et al. Targeting channelrhodopsin-2 to ON-bipolar cells with vitreally administered AAV Restores ON and OFF visual responses in blind mice. Mol Ther. 23(1): 7-16 (2015).
15. Jan, L.Y. and Jan, Y. N. L-Glutamate as an excitatory transmitter at the Drosophila larval neuromuscular junction . J. Physiol. 262, 215-236 (1976)
16. Kato, H. E. et al. Crystal structure of the channelrhodopsin light-gated cation channel. Nature. 428, 369-374 (2012)
Claims (15)
- 配列番号1(ChR-2)の全長配列に対し少なくとも84%の類似性及び/又は少なくとも75%の同一性を有するアミノ酸配列を含む変異型光誘導性イオンチャネルであって、
該変異型光誘導性イオンチャネルは、親光誘導性イオンチャネルと、配列番号1のF219に相当する位置における置換のみ異なっており、
前記置換は、-60 mVのクランプ電位、140 mM NaCl、2mM CaCl2、2MgCl2、10 mM HEPES、pH 7.4のバス溶液、及び110 mM NaCl、2 mM MgCl2、10 mM EGTA、10 mM HEPES、pH 7.4のピペット溶液で、全細胞構成におけるパッチクランプ測定により比較したとき、前記親チャネルと比べて、前記変異型チャネルのオフキネティクスを加速する、前記変異型光誘導性イオンチャネル。 - 前記変異型光誘導性イオンチャネルは、配列番号1(ChR-2)の全長に対し、少なくとも86%、好ましくは少なくとも88%、より好ましくは少なくとも90%、より好ましくは少なくとも92%、より好ましくは少なくとも94%、より好ましくは少なくとも96%、より好ましくは少なくとも98%、より好ましくは少なくとも99%の類似性を有する;及び/又は
前記変異型光誘導性イオンチャネルは、配列番号1(ChR-2)の全長に対し、少なくとも80%、好ましくは少なくとも85%、より好ましくは少なくとも90%、より好ましくは少なくとも95%、より好ましくは少なくとも96%、より好ましくは少なくとも97%、より好ましくは少なくとも98%、より好ましくは少なくとも99%の同一性を有する、
請求項1に記載の変異型光誘導性イオンチャネル。 - 前記置換はF219Yである、請求項1又は2に記載の変異型光誘導性イオンチャネル。
- 前記変異型チャネルは、配列番号4:
Cys-Arg-Xaa3-Xaa4-Val-Xaa6-Xaa7-Met-Ala-Trp-Xaa11-Tyr-Phe-Val-Xaa15-Trp-Gly-Met-Phe-Pro-Xaa21-Leu-Phe-Xaa24-Leu
のモチーフ、
ここでXaa3はGln又はGluである、好ましくはここでXaa3はGlnである;
ここでXaa4はVal又はLeuである、好ましくはここでXaa4はValである;
ここでXaa6はThr又はArgである、好ましくはここでXaa6はThrである;
ここでXaa7はGly、Val又はAlaである、好ましくはここでXaa7はGlyである;
ここでXaa11はLeu又はThrである、好ましくはここでXaa11はLeuである;
ここでXaa15はSer又はAlaである、好ましくはここでXaa15はSerである;
ここでXaa21はIle又はValである、好ましくはここでXaa21はIleである;および
ここでXaa24はIle又はLeuである、好ましくはここでXaa24はIleである、
を含む、請求項3に記載の変異型光誘導性イオンチャネル。 - 前記変異型チャネルは、配列番号1の位置L132に相当する位置においてCys、Ser、Glu、Asp、又はThrを更に含む、特にここで、前記変異型チャネルは、配列番号1の位置L132に相当する位置においてCysを含む、請求項1〜4のいずれか1項に記載の変異型光誘導性イオンチャネル。
- 前記変異型光誘導性イオンチャネルは、配列番号1(ChR-2)のアミノ酸配列を含む、好ましくは配列番号1(ChR-2)のアミノ酸配列からなる、但し、位置F219における前記置換及び場合により配列番号1の位置132におけるアミノ酸を除く;又は
前記変異型光誘導性イオンチャネルは、配列番号2(VChR1)のアミノ酸配列を含む、好ましくは配列番号2(VChR1)のアミノ酸配列からなる、但し、位置F214における前記置換及び場合により配列番号1のL132に相当する配列番号2の位置におけるアミノ酸を除く;又は
前記変異型光誘導性イオンチャネルは、配列番号3(ReaChR)のアミノ酸配列を含む、好ましくは配列番号3(ReaChR)のアミノ酸配列からなる、但し、位置F259における前記置換及び場合により配列番号1のL132に相当する配列番号3の位置におけるアミノ酸を除く、
請求項1〜5のいずれか1項に記載の変異型光誘導性イオンチャネル。 - 前記光誘導性イオンチャネルは、以下のアミノ酸残基:配列番号1の位置253に相当する位置におけるアスパラギン酸;配列番号1の位置257に相当する位置におけるリジン;配列番号1の位置260に相当する位置におけるトリプトファン;配列番号1の位置123に相当する位置におけるグルタミン酸;配列番号1の位置134に相当する位置におけるヒスチジン又はアルギニン、好ましくはアルギニン;配列番号1の位置128に相当する位置におけるスレオニン、セリン、又はアラニン;及び/又は配列番号1の位置156に相当する位置におけるアラニン;の少なくとも1つを追加的に含む、請求項1〜6のいずれか1項に記載の変異型光誘導性イオンチャネル。
- 請求項1〜7のいずれか1項に記載の変異型光誘導性イオンチャネルをコードするヌクレオチド配列を含む核酸構築物。
- 請求項1〜7のいずれか1項に記載の光誘導性イオンチャネルをコードするヌクレオチド配列又は請求項8に記載の核酸構築物を含む発現ベクター。
- 請求項8に記載の核酸構築物又は請求項9に記載の発現ベクターを含む細胞。
- 前記細胞は哺乳類細胞であり、好ましくはここで前記細胞は、
(a)海馬細胞、光受容体細胞、網膜桿体細胞、網膜錐体細胞、網膜神経節細胞、双極神経細胞、神経節細胞、偽単極性神経細胞、多極神経細胞、錐体神経細胞、プルキンエ細胞、又は顆粒細胞;あるいは、
(b)神経芽腫細胞、特にNG108-15、HEK293細胞;COS細胞;BHK細胞;CHO細胞;骨髄腫細胞;又はMDCK細胞である、請求項10に記載の細胞。 - ハイスループットスクリーニングにおける、請求項1〜7のいずれか1項に記載の光誘導性イオンチャネルあるいは請求項10又は11に記載の細胞の使用。
- 神経細胞の光刺激のための、請求項1〜7のいずれか1項に記載の光誘導性イオンチャネルの非治療的使用。
- 医薬における使用のための請求項1〜7のいずれか1項に記載の光誘導性イオンチャネル。
- 請求項1〜7のいずれか1項に記載の光誘導性イオンチャネル、請求項8に記載の核酸構築物、請求項9に記載の発現ベクター、あるいは請求項10又は11に記載の細胞、を含む非ヒト動物。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP16172992 | 2016-06-03 | ||
EP16172992.6 | 2016-06-03 | ||
PCT/EP2017/063425 WO2017207745A1 (en) | 2016-06-03 | 2017-06-02 | Mutant light-inducible ion channel of channelrhodopsin |
Publications (1)
Publication Number | Publication Date |
---|---|
JP2019522469A true JP2019522469A (ja) | 2019-08-15 |
Family
ID=56132766
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2018563484A Pending JP2019522469A (ja) | 2016-06-03 | 2017-06-02 | チャネルロドプシンの変異型光誘導性イオンチャネル |
Country Status (6)
Country | Link |
---|---|
US (1) | US20190218256A1 (ja) |
EP (1) | EP3464341A1 (ja) |
JP (1) | JP2019522469A (ja) |
KR (1) | KR20190020702A (ja) |
CN (1) | CN109476720A (ja) |
WO (1) | WO2017207745A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP4223768A1 (en) | 2022-02-04 | 2023-08-09 | Georg-August-Universität Göttingen Stiftung Öffentlichen Rechts, Universitätsmedizin | Novel mutant bacteriorhodopsin-like-channelrhodopsin ion channel |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012032103A1 (en) * | 2010-09-08 | 2012-03-15 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Mutant channelrhodopsin 2 |
US8202699B2 (en) * | 2002-04-11 | 2012-06-19 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Use of biological photoreceptors as directly light-activated ion channels |
US8759492B2 (en) * | 2011-08-17 | 2014-06-24 | The Regents Of The University Of California | Engineered red-shifted channelrhodopsin variants |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009119782A1 (ja) * | 2008-03-24 | 2009-10-01 | 国立大学法人東北大学 | 改変された光受容体チャネル型ロドプシンタンパク質 |
EA201001621A1 (ru) * | 2008-04-18 | 2011-06-30 | Новартис Форшунгсштифтунг, Цвайгнидерлассунг Фридрих Мишер Инститьют Фор Байомедикал Рисёрч | Новые терапевтические средства и способы лечения слепоты |
DK3214094T3 (da) | 2011-11-12 | 2020-07-13 | Massachusetts Inst Technology | Kanalrhodopsiner til optisk kontrol af celler |
HUE040487T2 (hu) * | 2012-03-05 | 2019-03-28 | Univ Wayne State | Csatornarodopszin-2 (CHOP2) mutációk azonosítása és alkalmazási eljárások |
-
2017
- 2017-06-02 US US16/306,673 patent/US20190218256A1/en not_active Abandoned
- 2017-06-02 JP JP2018563484A patent/JP2019522469A/ja active Pending
- 2017-06-02 KR KR1020187037967A patent/KR20190020702A/ko not_active Application Discontinuation
- 2017-06-02 WO PCT/EP2017/063425 patent/WO2017207745A1/en unknown
- 2017-06-02 CN CN201780034434.5A patent/CN109476720A/zh active Pending
- 2017-06-02 EP EP17728808.1A patent/EP3464341A1/en not_active Withdrawn
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8202699B2 (en) * | 2002-04-11 | 2012-06-19 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Use of biological photoreceptors as directly light-activated ion channels |
WO2012032103A1 (en) * | 2010-09-08 | 2012-03-15 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Mutant channelrhodopsin 2 |
US8759492B2 (en) * | 2011-08-17 | 2014-06-24 | The Regents Of The University Of California | Engineered red-shifted channelrhodopsin variants |
Non-Patent Citations (1)
Title |
---|
ZHANG, FENG ET AL."RED-SHIFTED OPTOGENETIC EXCITATION: A TOOL FOR FAST NEURAL CONTROL DERIVEDFROM V, JPN6021027687, 2008, ISSN: 0004982156 * |
Also Published As
Publication number | Publication date |
---|---|
EP3464341A1 (en) | 2019-04-10 |
US20190218256A1 (en) | 2019-07-18 |
CN109476720A (zh) | 2019-03-15 |
WO2017207745A1 (en) | 2017-12-07 |
KR20190020702A (ko) | 2019-03-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
RU2595384C2 (ru) | Мутантный канальный родопсин-2 | |
Lin et al. | Optogenetic inhibition of synaptic release with chromophore-assisted light inactivation (CALI) | |
CN105941328B (zh) | 一种鉴定抑制前额叶皮质中的兴奋性或抑制性神经元的去极化的化合物的系统 | |
JP5544659B2 (ja) | 改変された光受容体チャネル型ロドプシンタンパク質 | |
US20030040080A1 (en) | Bio-synthetic photostimulators and methods of use | |
US20120214188A1 (en) | Light-activated ion channel molecules and uses thereof | |
US10882892B2 (en) | Channelrhodopsin variants and uses thereof | |
US20170362281A1 (en) | Mutant nq-rhodopsin kr 2 | |
JP2019522469A (ja) | チャネルロドプシンの変異型光誘導性イオンチャネル | |
JP7175880B2 (ja) | Chrimsonの変異型光誘導性イオンチャネル | |
EP4223768A1 (en) | Novel mutant bacteriorhodopsin-like-channelrhodopsin ion channel | |
WO2021182646A1 (ja) | 色認識のための光応答性タンパク質及びその利用 | |
US11542314B2 (en) | Step-function channelrhodopsins for optical control of cells | |
JP2020516295A (ja) | 新規光遺伝学ツール |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20200602 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210727 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20211026 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220127 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20220531 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20220805 |
|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20230207 |