| Sequence-based reagent |
gRNA control |
This study |
5’-ATATTTCGGCAGTTGCAGCA-3'
|
CRISPR/Cas9Construction |
| Sequence-based reagent |
gRNA MDGA2 |
This study |
5’-ATTTAGTGTACGGTCTCGTG-3’
|
CRISPR/Cas9Construction |
| Sequence-based reagent |
gRNA MDGA1 |
This study |
5’-CTTCAACGTACGAGCCCGGG-3’
|
CRISPR/Cas9Construction |
| Sequence-based reagent |
AP tag sequence |
This study |
5´-GGCCTGAACGAtATCTTCGAGGCCCAG
AAGATCGAGTGGCACGAG-3´
|
AP tag plasmids |
| Sequence-based reagent |
MDGA1 CRISPR validation primers |
This study |
5´-GGGAAGAGGTAGAGACCCAAGT-3´
5´-CCTCCATCAACACATAACGAAA-3´
|
CRISPR/Cas9Validation |
| Sequence-based reagent |
MDGA2 CRISPR validation primers |
This study |
5´-GCTGATAGGGAAGGACAGACAG-3´
5´-TAAATCCAAGACTGCAAGAGCC-3´
|
CRISPR/Cas9validation |
| Sequence-based reagent |
MDGA1 RTqPCR primers |
This study |
5'-GTTCTACTGCTCCCTCAACC-3'
5'-CGTTACCTTTATTACCGCTGAG-3'
|
RTqPCR |
| Sequence-based reagent |
MDGA2 RTqPCR primers |
This study |
5´-AAGGTGACATCGCCATTGAC-3'5'-CCACGGAATTCTTAGTTGGTAGG-3´
|
RTqPCR |
| Sequence-based reagent |
U6 RTqPCR primers |
This study |
5′-GGAACGATACAGAGAAGATTAGC-3′
5′-AAATATGGAACGCTTCACGA-3′
|
RTqPCR |
| Sequence-based reagent |
SDHA RTqPCR primers |
This study |
5′-TGCGGAAGCACGGAAGGAGT-3′
5′-CTTCTGCTGGCCCTCGATGG-3′
|
RTqPCR |
| Sequence-based reagent |
Template switching oligo |
This study |
5′-AAGCAGTGGTATCAACGCAGAGTACrGrG + G-3′ |
Reverse transcription for RNA-seq |
| Genetic reagent |
pCMV6-XL4 |
Origene #pCMVXL4 |
|
MDGA1 expression |
| Recombinant DNA reagent |
V5-MDGA1(R.norvegicus) |
A.M. Craig (University of British Columbia, Canada) Pettem et al., 2013
|
|
Neuron electroporation |
| Recombinant DNA reagent |
V5-MDGA2(R.norvegicus) |
A.M. Craig (University of British Columbia, Canada) |
|
Neuron electroporation |
| Recombinant DNA reagent |
HA-MDGA1(R.norvegicus) |
A.M. Craig (University of British Columbia, Canada) Pettem et al., 2013
|
|
Neuron electroporation |
| Recombinant DNA reagent |
HA-MDGA2(R.norvegicus) |
A.M. Craig (University of British Columbia, Canada) Pettem et al., 2013
|
|
Neuron electroporation |
| Recombinant DNA reagent |
shMDGA1(R.norvegicus) |
A.M. Craig (University of British Columbia, Canada) Pettem et al., 2013
|
|
Neuron electroporation |
| Recombinant DNA reagent |
shMORB(R.norvegicus) |
A.M. Craig (University of British Columbia, Canada) Pettem et al., 2013
|
|
Neuron electroporation |
| Recombinant DNA reagent |
HA-MDGA1 rescue(R.norvegicus) |
A.M. Craig (University of British Columbia, Canada) Pettem et al., 2013
|
|
Neuron electroporation |
| Recombinant DNA reagent |
AP-NLGN1(M. musculus) |
A.Ting (Stanford University, USA) |
|
Neuron electroporation |
| Recombinant DNA reagent |
BirAER(M. musculus) |
A.Ting (Stanford University, USA) |
|
Neuron electroporation |
| Recombinant DNA reagent |
mApple.V5-MDGA2 rescue(R.norvegicus) |
A.Ting (Stanford University, USA) Loh et al., 2016
|
|
Plasmid construction |
| Recombinant DNA reagent |
shMDGA2(R.norvegicus) |
A.Ting (Stanford University, USA) Loh et al., 2016
|
|
Neuron electroporation |
| Recombinant DNA reagent |
AP-MDGA1 rescue(R.norvegicus) |
This paper |
|
Obtained with HD-In-Fusion kit |
| Recombinant DNA reagent |
AP-MDGA2 rescue(R.norvegicus) |
This paper |
|
Obtained with HD-In-Fusion kit |
| Recombinant DNA reagent |
pSpCas9(BB)–2A-GFP (PX458) |
Adgene #48,138 |
RRID:Adgene_48138
|
|
| Recombinant DNA reagent |
shNLGN1(M. musculus) |
P.Scheiffele (Biozentrum, Basel) |
|
Neuron electroporation |
| Recombinant DNA reagent |
HA-NLGN1(M. musculus) |
P.Scheiffele (Biozentrum, Basel) |
|
Obtained with HD-In-Fusion kit |
| Recombinant DNA reagent |
AP-NLGN1 rescue(M. musculus) |
Chamma et al., 2016a
|
|
Neuron electroporation |
| Recombinant DNA reagent |
Homer1c-DsRed |
Mondin et al., 2011
|
|
Neuron electroporation |
| Recombinant DNA reagent |
GFP-NRXN1β |
M.Missler (Münster University, Germany) Neupert et al., 2015
|
|
Neuron electroporation |
| Recombinant DNA reagent |
GFP-GPI |
Renner et al., 2009
|
|
Neuron electroporation |
| Recombinant DNA reagent |
pCAG_Xph20-eGFP-CCR5TC |
|
RRID:Adgene_135530
|
Neuron electroporation |
| Recombinant DNA reagent |
pCAG_Xph20-mRuby2_CCR5TC |
|
RRID:Adgene_135531
|
Neuron electroporation |
| Peptide, recombinant protein |
Anti GFP nanobody |
Chamma et al., 2016a
|
|
GFP labelling uPAINT, dSTORM |
| Peptide, recombinant protein |
mSA |
Chamma et al., 2017
|
|
AP-biotin labelling uPAINT, dSTORM |
| Cell line |
COS-7 from ECACC |
Sigma-AldrichAcc Nc 87021302Lot 15I032 passage + 4 |
RRID:CVCL_0224
|
Cell surface cluster assays, protein pull-down |
| Cell line |
HEK-293T from ECACC |
Sigma-AldrichAcc Nc 12022001Lot 16G020 Passage + 6 |
RRID:CVCL_0063
|
MDGA1 peptide production |
| Chemical compound, drug |
Bicuculine |
TOCRIS #0130/50 |
RRID:SCR_003689
|
Block inhibitory synaptic transmission |
| Chemical compound, drug |
Phosphocreatine |
Calcbiochem#2380–5 GM |
|
RNA extraction |
| Chemical compound, drug |
Atto 647 N |
Atto-Tec |
|
Coupled to anti GluA2 antibody |
| Chemical compound, drug |
STAR 635 P |
Abberior |
|
Coupled to mSA |
| Chemical compound, drug |
Alexa Fluor 647 |
Thermo Fischer Scientific |
|
Coupled to mSA |
| Chemical compound, drug |
V5 tag Fab fragment |
Abnova #RAB00032
|
|
Coupled to flurophores |
| Chemical compound, drug |
QIAzol Lysis Reagent |
Qiazol |
|
RTqPCR |
| Chemical compound, drug |
X- tremeGENE |
Transfection Reagent, Roche #6366546001 |
RRID:SCR_001326
|
COS7 cells transfection |
| Chemical compound, drug |
Ribolock |
Thermo Fisher Scientific #E00381
|
|
RNase inhibitor |
| Commercial assay, kit |
In-Fusion HD Cloning Kit |
Takara Bio#639,642 |
|
Plasmid construction |
| Commercial assay, kit |
MycoAlert Mycoplasma Detection Kit |
Lonza# LT07-218 |
Lot number: 0000312202 |
Mycoplasma detection in cell lines |
| Commercial assay, kit |
T7 endonuclease based method |
GeneArt Genomic detection kit, Thermo Fisher Scientific #A24372
|
|
CRISPR validation |
| Commercial assay, kit |
Direct-Zol RNA microprep |
Zymo Research cat#R2062 |
|
RTqPCR |
| Commercial assay, kit |
Maxima First Strand cDNA Synthesis kit |
Thermo Fischer Scientific # K1641 |
|
RTqPCR |
| Commercial assay, kit |
Dynabeads Protein G |
Thermo Fisher Scientific #11,004D |
RRID:SCR_008452
|
NLGN pull down |
| Commercial assay, kit |
Dynabeads M-280 |
Thermo Fisher Scientific #11,205D |
|
NLGN pull down |
| Commercial assay, kit |
ClarityWestern blot ECL |
Bio-Rad#170–5061 |
|
|
| Antibody |
Actin (mouse monoclonal) |
Sigma-Aldrich#A5316 |
RRID:AB_476743
|
(1:10,000 WB) |
| Antibody |
βIII tubulin (rabbit polyclonal) |
Abcam#18,207 |
RRID:AB_444319
|
(1:25,000 WB) |
| Antibody |
GFP (mouse monoclonal) |
Sigma-Aldrich#11814460001 |
RRID:AB_390913
|
(1:1000 WB) |
| Antibody |
HA (rat monoclonal) |
Roche#11867423001 |
RRID:AB_390918
|
(1:1000 WB) |
| Antibody |
HA (rabbit monoclonal) |
Cell Signaling #3,724 |
RRID:AB_10693385
|
|
| Antibody |
MDGA1 (rabbit polyclonal) |
Synaptic Systems#421,002 |
RRID:AB_2800520
|
1:50 ICC(1:500 WB) |
| Antibody |
GluA1 (rabbit polyclonal) |
Agrobio (clone G02141) |
- |
(1:50 ICC) |
| Antibody |
GluA2 (mouse monoclonal) |
E.Gouaux (OSHU, Vollum Institute, Portland) |
- |
(1:200 ICC) |
| Antibody |
NLGN1/2/3/4 (rabbit polyclonal) |
Synaptic Systems#129,213 |
RRID:AB_2619812
|
NLGN pull down (1:1000 WB) |
| Antibody |
PSD-95 (mouse monoclonal) |
Thermo Fischer Scientific #MA1-046 |
RRID:AB_2092361
|
1:200 ICC(1:2000 WB) |
| Antibody |
Gephyrin (mouse monoclonal) |
Synaptic Systems#147,111 |
RRID:AB_887719
|
(1:2000 ICC) |
| Antibody |
VGLUT1 (guinea pig polyclonal) |
Merck Millipore#AB5905 |
RRID:AB_2301751
|
(1:2000 ICC) |
| Antibody |
VGAT (guinea pig polyclonal) |
Synaptic Systems#131,004 |
RRID:AB_887873
|
(1:1,000 ICC) |
| Antibody |
Synaptophysin (mouse monoclonal) |
Sigma-Aldrich# S5768 |
Clone SVP-38RRID:AB_477523
|
(1:2000 WB) |
| Antibody |
Tubulin (mouse monoclonal) |
Sigma-Aldrich#T4026 |
RRID:AB_477577
|
(1:5000 WB) |
| Antibody |
MAP-2 (mouse monoclonal) |
Sigma-Aldrich#M4403 |
RRID:AB_477193
|
(1:2000 ICC) |
| Antibody |
p-tyrosine (mouse monoclonal) |
Cell Signaling#9,411 |
RRID:AB_331228
|
(1:1000 WB) |
| Antibody |
Anti-mouse HRP (donkey polyclonal) |
Jackson Immunoresearch #715-035-150 |
RRID:AB_2340770
|
(1:5000 WB) |
| Antibody |
Anti-mouse IRDye680LT(goat polyclonal) |
LICOR #926–68020 |
RRID:AB_10706161
|
(1:10,000 WB) |
| Antibody |
Anti-mouseIRDye 800LT(goat polyclonal) |
LICOR #926–32210 |
RRID:AB_26218442
|
(1:10,000 WB) |
| Antibody |
Anti-rabbit HRP (donkey polyclonal) |
Jackson Immunoresearch #711-035-152 |
RRID:AB_10015282
|
(1:5000 WB) |
| Antibody |
Anti-rabbitIRDye 680LT(goat polyclonal) |
LICOR #926–68021 |
RRID:AB_10706309
|
(1:10,000 WB) |
| Antibody |
Anti-rabbit IRDye800LT(goat polyclonal) |
LICOR #926–32211 |
RRID:AB_621843
|
(1:10,000 WB) |
| Antibody |
Anti-mouse Alexa488 (goat polyclonal) |
Thermo Scientific #A11001 |
RRID:AB_2534069
|
(1:1,000 ICC) |
| Antibody |
Anti-mouse Alexa 568 (goat polyclonal) |
Thermo Scientific #A11031 |
RRID:AB_144696
|
(1:1,000 ICC) |
| Antibody |
Anti-mouse Alexa 647 (goat polyclonal) |
Thermo Scientific #A21235 |
RRID:AB_2535804
|
(1:1,000 ICC) |
| Antibody |
Anti-rabbit Alexa488 (goat polyclonal) |
Thermo Scientific #A11008 |
RRID:AB_143165
|
(1:1,000 ICC) |
| Antibody |
Anti-rabbit Alexa 568 (goat polyclonal) |
Thermo Scientific #A11011 |
RRID:AB_143157
|
(1:1,000 ICC) |
| Antibody |
Anti-rabbit Alexa 647 (goat polyclonal) |
Thermo Scientific #A31576
|
RRID:AB_10374303
|
(1:1,000 ICC) |
| Antibody |
Anti-rabbit Alexa 555 (donkey polyclonal) |
Invitrogen#A32794 |
RRID:AB_2762834
|
(1:1,000 ICC) |
| Antibody |
Anti-rat Alexa 488 (goat polyclonal) |
Thermo Scientific #A11073 |
RRID:AB_2534117
|
(1:1,000 ICC) |
| Antibody |
Anti-rat Alexa 568 (goat polyclonal) |
Thermo Scientific #A11077 |
RRID:AB_2534121
|
(1:1,000 ICC) |
| Antibody |
Anti Rat Alexa 647 (goat polyclonal) |
Thermo Scientific #A21247 |
RRID:AB_141778
|
(1:1,000 ICC) |
| Antibody |
Anti guinea pig DyLight 405 (goat polyclonal) |
Jackson ImmunoResearch #106475003 |
RRID:AB_2337432
|
(1:1,000 ICC) |
| Antibody |
Anti-guinea pig Alexa488 (goat polyclonal) |
Thermo Scientific #A11008 |
RRID:AB_143165
|
(1:1,000 ICC) |
| Antibody |
Anti-guinea pig Alexa 568 (goat polyclonal) |
Thermo Scientific #A11075 |
RRID:AB_141954
|
(1:1,000 ICC) |
| Antibody |
Anti guinea pig Alexa 647 (goat polyclonal) |
Thermo Scientific #A21450 |
RRID:AB_2141882
|
(1:1,000 ICC) |
| Software, algorithm |
Metamorph |
Molecular devices |
RRID:SCR_002368
|
Image analysis |
| Software, algorithm |
Graphpad |
PRISM |
RRID:SCR_002798
|
Statistical analysis |
| Software, algorithm |
ChopChop |
Labun et al., 2019
|
|
CRISPR/Cas9 design |
| Software, algorithm |
ImageJ |
National Center for Microscopy and Imaging Research |
RRID:SCR_001935
|
Western-blot quantitation |
| Software, algorithm |
cutadapt |
Martin, 2011
|
|
RNA-seq |
| Software, algorithm |
fastp |
Chen et al., 2018
|
|
RNA-seq |
| Software, algorithm |
STAR |
Dobin et al., 2013
|
|
RNA-seq |
| Software, algorithm |
DESeq2 |
Love et al., 2014
|
|
RNA-seq |
