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技术与服务

光遗传

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:

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;

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;

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;

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;

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;

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460nm PT-1431 rAAV-hSyn-CheRiff-eGFP-WPRE-bGH polyA hSyn EGFP

hChR2

(H134R)

激发光波长473nm 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473nm
高频光激活适用
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473nm
高频光激活适用
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.), 590(inact.) PT-1430 rAAV-CaMKIIa-hChR2(C128S/D156A)-mCherry-WPRE-hGH polyA CaMKIIα mCherry
ChETA
激发光波长490nm
高频光激活适用
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470nm PT-1435 rAAV-hSyn-Chronos-EGFP-WPRE-bGH polyA hSyn EGFP
PT-1434 rAAV-hSyn-Chronos-tdTomato-WPRE-bGH polyA hSyn tdTomato

C1V1E122T-E162T)

激发光波长535nm 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590nm 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590-630nm 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475nm
单次光抑制长效应
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515nm 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589nm 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590nm 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566nm 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566nm 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632nm 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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