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光遗传技术服务
原理及应用介绍
光遗传学(Optogenetics)是指结合光学与遗传学手段,精确控制特定神经元活动的技术。该技术利用分子生物学、病毒生物学等手段,将外源光敏感蛋白基因导入活细胞中,在细胞膜结构上表达了光敏感通道蛋白;然后通过特定波长光的照射,控制细胞膜结构上的光敏感通道蛋白的激活与关闭;光敏感蛋白的激活和关闭可控制细胞膜上离子通道的打开与关闭,进而改变细胞膜电压的变化,如膜的去极化与超极化。当膜电压去极化超过一定阈值时就会诱发神经元产生可传导的电信号,即神经元的激活;相反,当膜电压超极化到一定水平时,就会抑制神经元动作电位的产生,即神经元的抑制。
实施方法
光遗传学技术主要包括病毒包装、病毒注射、光纤埋植、光调控、行为测试和电生理记录6个步骤。具体实施应该包括下面6个方面:
①需要寻找合适的光敏蛋白:例如起兴奋神经元作用的ChR2,起抑制神经元作的NpHR和Arch等;
②将遗传信息传递给靶细胞:一般通过病毒转导、转染、转基因动物等方式将光敏蛋白的遗传信息传递给靶细胞,依赖重组酶Cre的嗜神经病毒选择性表达光感基因;
③可控性演示:即通过导入光纤、控制激光来实现对神经元活动的精确控制;
④根据病毒的表达时间,通过特定频率的激光调控光敏感通道的打开与关闭;
⑤实验方法的有效性验证:一般采用电极记录神经元细胞膜内外电压变化,以此来验证光遗传的有效性;
⑥表型检测:通过行为学测试来评估神经元活动对动物行为的影响。
图1.光遗传技术示意图
图2.光纤埋置
图3.小鼠光遗传实验示意图
我司提供光遗传病毒及相关实验外包服务
表1 光遗传病毒现货列表
名称 |
功能描述 |
产品编号 |
产品名称 |
启动子 |
Cre/Flp依赖 |
标签 |
---|---|---|---|---|---|---|
光激活 |
||||||
CheRiff |
激发光波长 |
PT-1431 |
rAAV-hSyn-CheRiff-eGFP-WPRE-bGH polyA |
hSyn |
|
EGFP |
hChR2 |
激发光波长 |
PT-0296 |
PFD-rAAV-CaMKIIa-hChR2(H134R)-EYFP-WPRE-hGH polyA |
CaMKIIα |
|
EYFP |
PT-0297 |
PFD-rAAV-CaMKIIa-hChR2(H134R)-mCherry-WPRE-hGH polyA |
CaMKIIα |
|
mCherry |
||
PT-1317 |
rAAV-hSyn-hChR2(H134R)-EYFP-WPRE-hGH polyA |
hSyn |
|
EYFP |
||
PT-0150 |
rAAV-hSyn-hChR2(H134R)-mCherry-WPRE-hGH polyA |
hSyn |
|
mCherry |
||
PT-0158 |
rAAV-EF1a-ChR2-EYFP-WPRE-hGH polyA |
EF1α |
|
EYFP |
||
PT-0992 |
rAAV-CAG-hChR2(H134R)-mCherry-WPRE-hGH polyA |
CAG |
|
mCherry |
||
PT-0985 |
PFD-AAV-hSyn-DIO-hChR2(H134R)-EYFP-WPRE-hGH polyA |
hSyn |
Cre依赖 |
EYFP |
||
PT-0001 |
rAAV-EF1α-DIO-hChR2(H134R)-EYFP-WPRE-hGH polyA |
EF1α |
Cre依赖 |
EYFP |
||
PT-0002 |
rAAV-EF1α-DIO-hChR2(H134R)-mCherry-WPRE-hGH polyA |
EF1α |
Cre依赖 |
mCherry |
||
PT-0474 |
rAAV-CAG-DIO-hChR2(H134R)-EYFP-WPREs |
CAG |
Cre依赖 |
EYFP |
||
PT-0189 |
rAAV-hSyn-fDIO-hChR2(H134R)-mCherry-WPRE-hGH polyA |
hSyn |
Flp依赖 |
EGFP |
||
PT-1375 |
rAAV-CAG-hChR2(H134R)-EYFP-WPRE-hGH polyA |
CAG |
|
EYFP |
||
PT-0235 |
rAAV-CAG-hChR2(H134R)-tdTomato-WPRE-SV40 polyA |
CAG |
|
tdTomato |
||
PT-0067 |
PFD-rAAV-hSyn-Coff Fon-hChR2(H134R)-EYFP-WPRE-hGH polyA |
hSyn |
Creoff/Flpon |
EYFP |
||
PT-0066 |
rAAV-nEF1a-Con Foff-hChR2(H134R)-EYFP-WPRE-hGH polyA |
nEF1α |
Creon/Flpoff |
EYFP |
||
PT-0065 |
rAAV-hSyn-Con Fon-hChR2(H134R)-EYFP-WPRE-hGH polyA |
hSyn |
Creon/Flpon |
EYFP |
||
PT-0968 |
rAAV-EF1α-Con Fon hChR2(H134R)-EYFP-WPRE-hGH polyA |
EF1α |
Creon/Flpon |
EYFP |
||
hChR2 (E123T/T159C) |
激发光波长 |
PT-0004 |
rAAV-CaMKIIa-hChR2(E123T/T159C)-EYFP-WPRE-hGH polyA |
CaMKIIα |
|
EYFP |
PT-0005 |
rAAV-CaMKIIa-hChR2(E123T/T159C)-mCherry-WPRE-hGH polyA |
CaMKIIα |
|
mCherry |
||
PT-0807 |
rAAV-hSyn-hChR2(E123T/T159C)-EYFP-WPRE-hGH polyA |
hSyn |
|
EYFP |
||
PT-0491 |
rAAV-hSyn-hChR2(E123T/T159C)-mCherry-WPRE-hGH polyA |
hSyn |
|
mCherry |
||
PT-0014 |
rAAV-EF1α-DIO-hChR2(E123T/T159C)-EYFP-WPRE-hGH polyA |
EF1α |
Cre依赖 |
EYFP |
||
PT-0015 |
rAAV-EF1α-DIO-hChR2(E123T/T159C)-mCherry-WPRE-hGH polyA |
EF1α |
Cre依赖 |
mCherry |
||
OChIEF (E163A/T199C) |
激发光波长 |
PT-1598 |
rAAV-CaMKIIa-OChIEF(E163A T199C)-mCherry-WPRE-hGH polyA |
CaMKIIα |
|
mCherry |
PT-1594 |
rAAV-CaMKIIa-OChIEF(E163A T199C)-P2A-mCherry-WPRE-hGH polyA |
CaMKIIα |
|
mCherry |
||
PT-1182 |
rAAV-CaMKIIa-DIO-oChIEF-P2A-dTomato-WPRE-bGH polyA |
CaMKIIα |
Cre依赖 |
dTomato |
||
PT-0228 |
rAAV-EF1a-DIO-oChIEF(E163A/T199C)-P2A-dTomato-WPRE-bGH polyA |
EF1α |
Cre依赖 |
mCherry |
||
SSFO |
470(activ.), |
PT-1430 |
rAAV-CaMKIIa-hChR2(C128S/D156A)-mCherry-WPRE-hGH polyA |
CaMKIIα |
|
mCherry |
ChETA |
激发光波长 |
PT-0559 |
PFD-rAAV-hSyn-ChETA-EGFP-WPRE-hGH polyA |
hSyn |
|
EGFP |
PT-0232 |
rAAV-EF1a-DIO-ChETA-EYFP-WPRE-hGH polyA |
EF1α |
Cre依赖 |
EYFP |
||
Chronos |
激发光波长 |
PT-1435 |
rAAV-hSyn-Chronos-EGFP-WPRE-bGH polyA |
hSyn |
|
EGFP |
PT-1434 |
rAAV-hSyn-Chronos-tdTomato-WPRE-bGH polyA |
hSyn |
|
tdTomato |
||
C1V1(E122T-E162T) |
激发光波长 |
PT-0972 |
rAAV-CaMKIIa-C1V1-EGFP-WPRE-hGH polyA |
CaMKIIα |
|
EGFP |
PT-1429 |
rAAV-CaMKIIa-C1V1 (t/t)-TS-EYFP-WPRE-hGH polyA |
CaMKIIα |
|
EYFP |
||
ChrimsonR |
激发光波长 |
PT-1433 |
rAAV-hSyn-ChrimsonR-EGFP-WPRE-hGH polyA |
hSyn |
|
EGFP |
PT-1432 |
rAAV-hSyn-ChrimsonR-tdTomato-WPRE-bGH polyA |
hSyn |
|
tdTomato |
||
PT-1374 |
rAAV-hSyn-DIO-ChrimsonR-mCherry-WPRE-hGH polyA |
hSyn |
Cre依赖 |
mCherry |
||
ReachR |
激发光波长 |
PT-0063 |
rAAV-CaMKIIa-ReachR-citrine-WPRE-hGH polyA |
CaMKIIα |
|
citrine |
PT-0064 |
rAAV-EF1α-DIO-ReachR-citrine-WPRE-hGH polyA |
EF1α |
Cre依赖 |
citrine |
||
光抑制 |
||||||
SwiChRca |
激发光波长 |
PT-1437 |
rAAV-CaMKIIa-SwiChRca-TS-EYFP-WPRE-hGH polyA |
CaMKIIα |
|
EYFP |
PT-1438 |
rAAV-Ef1a-DIO-SwiChRca-TS-EYFP-WPRE-hGH polyA |
EF1α |
Cre依赖 |
EYFP |
||
GtACR1 |
激发光波长 |
PT-1156 |
rAAV-EF1a-GtACR1-P2A-EGFP-WPRE-hGH polyA |
EF1α |
|
EGFP |
PT-1368 |
rAAV-EF1a-DIO-GtACR1-P2A-EGFP-WPRE-hGH polyA |
EF1α |
Cre依赖 |
EGFP |
||
PT-1386 |
PFD-rAAV-nEF1a-fDIO-GtACR1-P2A-EGFP-WPRE-hGH polyA |
nEF1a |
Flp依赖 |
EGFP |
||
eNpHR3.0 |
激发光波长 |
PT-0008 |
rAAV-CaMKIIa-eNpHR3.0-EYFP-WPRE-hGH polyA |
CaMKIIα |
|
EYFP |
PT-0009 |
rAAV-CaMKIIa-eNpHR3.0-mCherry-WPRE-hGH polyA |
CaMKIIα |
|
mCherry |
||
PT-0010 |
rAAV-hSyn-eNpHR3.0-EYFP-WPRE-hGH polyA |
hSyn |
|
EYFP |
||
PT-0011 |
rAAV-hSyn-eNpHR3.0-mCherry-WPRE-hGH polyA |
hSyn |
|
mCherry |
||
PT-0006 |
rAAV-EF1α-DIO-eNpHR3.0-EYFP-WPRE-hGH polyA |
EF1α |
Cre依赖 |
EYFP |
||
PT-0007 |
rAAV-EF1α-DIO-eNpHR3.0-mCherry-WPRE-hGH polyA |
EF1α |
Cre依赖 |
mCherry |
||
PT-0689 |
PFD-rAAV-CAG-DIO-eNpHR3.0-EYFP-WPRE-pA |
CAG |
Cre依赖 |
EYFP |
||
PT-0724 |
rAAV-EF1a-eNpHR3.0-EYFP-WPRE-hGH polyA |
EF1a |
|
EYFP |
||
PT-1155 |
rAAV-hSyn-Coff Fon-eNpHR3.0-EYFP-WPRE-hGH polyA |
hSyn |
Creoff/Flpon |
EYFP |
||
PT-1153 |
rAAV-hSyn-Con Foff-eNpHR3.0-EYFP-WPRE-hGH polyA |
hSyn |
Creon/Flpoff |
EYFP |
||
PT-1154 |
rAAV-hSyn-Con Fon-eNpHR3.0-EYFP-WPRE-hGH polyA |
hSyn |
Creon/Flpon |
EYFP |
||
PT-1261 |
PFD-rAAV-nEF1a-fDIO-eNpHR3.0-mCherry-WPRE-hGH polyA |
nEF1a |
Flp依赖 |
mCherry |
||
FLlnChR |
激发光波长 |
PT-1252 |
PFD-rAAV-EF1a-DIO-FLlnChR-EYFP-WPRE-hGHpA |
EF1α |
Cre依赖 |
EYFP |
PT-1253 |
PFD-rAAV-EF1a-DIO-FLlnChR-mCherry-WPRE-hGHpA |
EF1α |
Cre依赖 |
mCherry |
||
Arch/ArchT |
激发光波长 |
PT-0633 |
rAAV-EF1a-DIO-ArCH-EGFP-WPRE-hGH polyA |
EF1α |
Cre依赖 |
EGFP |
PT-0555 |
rAAV-CAG-DIO-ARCH-EYFP-WPRE-hGH polyA |
CAG |
Cre依赖 |
EYFP |
||
Arch/ArchT |
激发光波长 |
PT-0560 |
PFD-rAAV-hSyn-ArchT-EGFP-WPRE-hGH polyA |
hSyn |
|
EGFP |
PT-0233 |
rAAV-CaMKIIa-ArchT-EGFP-WPRE-hGH polyA |
CaMKIIα |
|
EGFP |
||
PT-0243 |
rAAV-CAG-FLEX-ArchT-EGFP-WPRE-SV40 polyA |
CAG |
Cre依赖 |
EGFP |
||
Jaws |
激发光波长 |
PT-0121 |
rAAV-hSyn-Jaws-EGFP-ER2-WPRE-pA |
hSyn |
|
EGFP |
PT-0120 |
rAAV-CaMKIIa-Jaws-KGC-EGFP-ER2-WPRE-SV40 polyA |
CaMKIIα |
|
EGFP |
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