Antibodies |
|
Mouse anti-GFP (clones 7.1 and 13.1) |
Roche |
11814460001 |
Mouse monoclonal anti-GAPDH (clone 6C5) |
Millipore Sigma |
MAB374 |
Rabbit polyclonal anti-Gαolf |
Corvol et al., 2001 |
N/A |
|
Chemicals, Peptides, and Recombinant Proteins |
|
Dulbecco’s modified Eagle’s medium |
Thermo Fisher Scientific |
11965-092 |
Fetal bovine serum |
Millipore Sigma |
12303C |
Sodium pyruvate |
Thermo Fisher Scientific |
11360-070 |
MEM non-essential amino acids |
Thermo Fisher Scientific |
11140-050 |
Penicillin-streptomycin |
Thermo Fisher Scientific |
15140-122 |
Puromycin |
Thermo Fisher Scientific |
A1113803 |
Neurobasal-A medium |
Thermo Fisher Scientific |
10888-022 |
GlutaMAX supplement |
Thermo Fisher Scientific |
35050-061 |
B-27 Supplement |
Thermo Fisher Scientific |
17504044 |
Hanks’ balanced salt solution |
Thermo Fisher Scientific |
14175-095 |
Papain |
Worthington Biochemical |
LS003126 |
DNase I |
Thermo Fisher Scientific |
18047019 |
Poly-D-lysine hydrobromide |
Millipore Sigma |
P6407 |
Matrigel |
Corning |
356230 |
Dulbecco’s phosphate-buffered saline |
Millipore Sigma |
D8537 |
Hanks’ balanced salt solution |
Millipore Sigma |
55037C |
CO2-independent medium |
Thermo Fisher Scientific |
18045-088 |
cOmplete, EDTA-free protease inhibitor cocktail |
Millipore Sigma |
11873580001 |
Dynabeads Protein G |
Thermo Fisher Scientific |
10004D |
Dopamine hydrochloride |
Millipore Sigma |
H8502 |
|
Critical Commercial Assays |
|
GloSensor cAMP Reagent |
Promega |
E1290 |
Nano-Glo Luciferase Assay Substrate (furimazine) |
Promega |
N1120 |
NucleoSpin RNA Plus Kit |
Takara Bio |
740984.10 |
Titanium One-Step RT-PCR Kit |
Takara Bio |
639503 |
|
Experimental Models: Cell Lines |
|
Human: HEK293T/17 |
ATCC |
CRL11268 |
Human: GNAS knockout HEK293T/17 |
This study |
N/A |
|
Experimental Models: Organisms/Strains |
|
Mouse: CAMPER |
Muntean et al., 2018 |
N/A |
Mouse: Drd1-Cre |
JAX |
Tg(Drd1-cre)EY262Gsat |
Mouse: C57BL/6 |
CRL |
C57BL/6NCrl |
|
Oligonucleotides |
|
Primer: Gαolf forward: GATCGAGAAGCAGTTGCAGAAAGAG |
This study |
N/A |
Primer: Gαolf reverse: CTTTGTCCACTTGGAATCGTGTCTC |
This study |
N/A |
Primer: GAPDH forward: GTCTTCACCACCATGGAGAAGG |
This study |
N/A |
Primer: GAPDH reverse: GAAGGCCATGCCAGTGAGCTTC |
This study |
N/A |
|
Recombinant DNA |
|
Plasmid: GNAS CRISPR guide RNA pLentiCRISPR v2 |
This paper |
N/A |
Plasmid: Dopamine D1 receptor |
cDNA Resource Center |
DRD0100000 |
Plasmid: Gαolf |
cDNA Resource Center |
GNA0L00000 |
Plasmid: Gαolf P102_V104del |
This paper |
N/A |
Plasmid: Gαolf F133L |
Dos Santos et al., 2016 |
N/A |
Plasmid: Gαolf V137M |
This paper |
N/A |
Plasmid: Gαolf V146M |
This paper |
N/A |
Plasmid: Gαolf E155K |
This paper |
N/A |
Plasmid: Gαolf V172I |
This paper |
N/A |
Plasmid: Gαolf R188C |
This paper |
N/A |
Plasmid: Gαolf G213S |
This paper |
N/A |
Plasmid: Gαolf Q214L |
cDNA Resource Center |
GNA0L000C0 |
Plasmid: Gαolf V228F |
This paper |
N/A |
Plasmid: Gαolf V234I |
This paper |
N/A |
Plasmid: Gαolf S239N |
This paper |
N/A |
Plasmid: Gαolf A311T |
This paper |
N/A |
Plasmid: Gαolf R329W |
Masuho et al., 2016 |
N/A |
Plasmid: Gαolf A353T |
This paper |
N/A |
Plasmid: Gαolf V354A |
This paper |
N/A |
Plasmid: Venus 156-239-Gβ2 |
This paper |
N/A |
Plasmid: Gβ2 |
cDNA Resource Center |
GNB0200000 |
Plasmid: Venus 1-155-Gγ7 |
Lohmann et al., 2017 |
N/A |
Plasmid: Gγ7 |
cDNA Resource Center |
GNG0700000 |
Plasmid: Flag-tagged Ric-8B |
Von Dannecker et al., 2006 |
N/A |
Plasmid: PTX-S1 |
Raveh et al., 2010 |
N/A |
Plasmid: masGRK3ct-Nluc |
Masuho et al., 2015b |
N/A |
Plasmid: CalfluxVTN |
Himmelreich et al., 2017 |
N/A |
Plasmid: pGloSensor-22F cAMP |
Promega |
E2301 |
Plasmid: mTrpM3α2-C-GFP |
Ghosh et al., 2016 |
N/A |
Plasmid: masGRK3ct |
Kammermeier et al., 2000 |
N/A |
Plasmid: pmCherry-N1 Vector |
Takara Bio |
632523 |
|
Software and Algorithms |
|
R version 3.4.2 |
R Core Team |
https://www.r-project.org/ |
ImageJ |
National Institute of Health |
SCR_003070 |
GraphPad Prism 6 |
Graphpad Software |
SCR_002798 |
SigmaPlot 12.5 |
SYSTAT Software |
SCR_003210 |
PyMol |
Schrödinger |
SCR_000305 |
Clampfit 10.3 |
Molecular Devices |
SCR_011323 |
T-Coffee |
Notredame et al., 2000 |
SCR_011818 |
BoxShade |
|
SCR_007165 |
|
Other |
|
GPCRdb |
(Pándy-Szekeres et al., 2018
|
http://gpcrdb.org/ |
gnomAD |
Lek et al., 2016 |
http://gnomad.broadinstitute.org/ |
REVEL |
Ioannidis et al., 2016 |
https://sites.google.com/site/revelgenomics/downloads |
ANNOVAR |
Wang et al., 2010 |
http://annovar.openbioinformatics.org/en/latest/ |