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AAV GRAB sensors

Li Yulong’s lab from Peking University develops cutting edge research tools, namely advanced imaging probes (GRAB sensors), including published dopamine (2018, Cell), acetylcholine (2018, Nature Biotechnology), and norepinephrine (2019, Neuron), to untangle the complexity of nervous system in space and in time, and helps to understand the changes of neurotransmitters in specific diseases, thus providing a new path for future precision medicine and new drug development. The GRAB sensors could be expressed in specific cell-types and are able to detect dynamics of extracellular neurotransmitters

Li Lab has deposited sensor plasmids at BrainVTA for distribution to the research community, and all GRAB sensors (including the lastest ACh, DA, NE, 5-HT, Ado, ATP, VIP, CCK and eCB sensors) with AAV are available.
Please go to pre-made AAV center to search the published sensors, for un-published ones, please contact Li lab for permission fist!

Features of published sensors

Name	Neurotransmitter	Version	Color	Backbones(From human)	Affinity	Signal Response Amplitude	Dynamics	Downstream Signal Coupling	Reference
ACh2.0	Acetylcholine	First generation	Green	M8 receptor	EC50=20nM	ΔF/F=90-200%; τon=200ms	τon~200ms, τoff~800ms	Weak	[1]
ACh3.0	Acetylcholine	Second generation	Green	M8 receptor	EC50=20nM	ΔF/F=60-100%; τon=200ms	τon~112ms,τoff~580ms	Hardly	[2]
ACh3.0-mut	Acetylcholine	Control of Second generation	Green	M8 receptor (W210A mutation)	EC50=20nM	ΔF/F=-0.18%; τon=200ms	/	Hardly	[2]
DA1m	Dopamine	First generation	Red	D2 receptor	EC50=3-10nM	ΔF/F=60-100%; τon=60ms, τoff=700ms	τon~60ms, τoff~700ms	Hardly	[3]
DA1h	Dopamine	First generation	Red	D2 receptor	EC50=3-10nM	ΔF/F=80-150%; τon=60ms, τoff=250ms	τon~140ms, τoff~2500ms	Hardly	[3]
DA(D104A)	Dopamine	Control of First generation	Red	D2 receptor (D104A and S110R mutation)	EC50>500nM	No effect	/	[3]
DA2m/DA4m	Dopamine	Second generation	Green/Red	D2 receptor	EC50=3-10nM	ΔF/F=30-70%; τon=80ms, τoff=100ms	τon~40ms, τoff~1300ms	Little	[4]
DA2h/DA4h	Dopamine	Second generation	Green/Red	D2 receptor	EC50=3-10nM	ΔF/F=80-120%; τon=70ms, τoff=200ms	τon~50ms, τoff~7300ms	Little	[4]
DA(D104A2nd)	Dopamine	Control of Second generation	Green/Red	D2 receptor (D104A and S110R mutation)	EC50>500nM	No effect	/	[4]
αDA1m (αDA2.5m)	Dopamine	First generation	Red	D2 receptor	EC50=3-10nM	ΔF/F=150-200%; τon=30ms, τoff=80ms	τon~80ms, τoff~770ms	Little	[4]
αDA1h (αDA2.5h)	Dopamine	First generation	Red	D2 receptor	EC50=3-10nM	ΔF/F=100-150%; τon=30ms, τoff=200ms	τon~60ms, τoff~2150ms	Little	[4]
αDA(D104A2nd)	Dopamine	Control of Second generation	Red	D2 receptor (D104A and S110R mutation)	EC50>500nM	No effect	/	[4]
NE1m(β2.1)	Norepinephrine	First generation	Green	α2A receptor	EC50=120nM	ΔF/F=200%; τon=30ms, τoff=70ms	τon ~70ms, τoff~ 750ms	Uncoupled	[5]
NE1h(β2.2)	Norepinephrine	First generation	Green	α2A receptor	EC50=120nM	ΔF/F=130%; τon=30ms, τoff=200ms	τon ~30ms, τoff~ 2000ms	Uncoupled	[5]
NEmut	Norepinephrine	/	Green	α2A receptor	EC50~0uM S5.46A mutation	No effect	/	/	[5]
Ado1.0	Adenosine	Control	Green	α2A receptor	EC50~60nM	ΔF/F0~130%	τon~36ms, τoff~1890ms	Hardly	[6]
Ado1.0mut	Adenosine	/	Green	α2A receptor	EC50~0uM F168A mutation	No effect	/	/	[6]
5-HT1.0	Serotonin	Control	Green	5-HT2C receptor	EC50~22nM	ΔF/F0~250%	τon~0.2s, τoff~3.1s	Uncoupled	[7]
5-HTmut	Serotonin	/	Green	5-HT2C receptor	EC50~0uM D1343.32Q mutation	No effect	/	/	[7]

FAQ:

1. What do the abbreviation h/m/l/mut mean, and what about the number?
h/m/l is the abbreviation of high/median/low, which indicates the sensors' apparent affinity towards transmitter. Most of the GRAB sensors have transmitter-insensitive mutant for comparison with GRAB sensors to check signal specificity. The number represents the version of sensor, the higher version, the better performance.
2. How to use GRAB sensors?
GRAB sensors are genetically encoded sensors for neurotransmitters, similar to other genetically encoded sensors (e.g. GCaMP). They could be expressed in specific cell-types and are able to detect dynamics of extracellular neurotransmitters in cultured cell, brain slice and living animal (e.g. fly, zebrafish, mice, rat, monkey etc.). For the aspect of instruments, GRAB sensors could perform well in epifluorescence microscopy, confocal microscopy, 2P microscopy and fiber photometry recording.
3. Can the sensors be expressed in vivo for a long-term period?
Yes. We can record good signals from GRAB sensor after expression for 3 to 6 months in mice brain by AAV infection.
4. Can GRAB sensors detect excitability of neurons?
No. The change of fluorescent intensity only indicates the dynamics of corresponding neurotransmitter, which has no direct correlation with excitability of neurons. Other approach is needed for detection of neuron excitability.

Reference

Jing, M., et al. (2018). "A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies." Nat Biotechnol 36(8): 726-737.
Jing, M., et al. (2020). "An optimized acetylcholine sensor for monitoring in vivo cholinergic activity." Nat Methods 17(11): 1139-1146.
Sun, F., et al. (2018). "A Genetically Encoded Fluorescent Sensor Enables Rapid and Specific Detection of Dopamine in Flies, Fish, and Mice." Cell 174(2): 481-496 e419.
Sun, F., et al. (2020). "Next-generation GRAB sensors for monitoring dopaminergic activity in vivo." Nat Methods 17(11): 1156-1166.
Feng, J., et al. (2019). "A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo Detection of Norepinephrine." Neuron 102(4): 745-761 e748.
Peng, W., et al. (2020). "Regulation of sleep homeostasis mediator adenosine by basal forebrain glutamatergic neurons." Science 369(6508).
Wan, J., et al. (2021). "A genetically encoded sensor for measuring serotonin dynamics." Nat Neurosci.

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