Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) |
CRACR2A | NCBI | Gene ID: 84,766 | |
| Strain, strain background (Escherichia coli) | DH5α | Thermo fisher Scientific | Cat# 18265017 | |
| Cell line (Homo sapiens) | Jurkat E6-1 T cells | ATCC | Not tested for mycoplasma | Cat# TIB-152 |
| Cell line (Homo sapiens) | HEK293T | ATCC | Not tested for mycoplasma | Cat# CRL-3216 |
| Cell line (Mus musculus) | CD40L-expressing L cell fibroblasts | Diehl et al., 2008 | Irradiated at 50 Gy for 50 minutes prior to use |
|
| Cell line (Mus musculus) | M2-10B4 bone marrow stromal cells | Lemoine et al., 1988 | Irradiated at 57 Gy for 57 minutes prior to use |
|
| Transfected construct (Homo sapiens) | Primers for plasmid construction | This paper | Supplementary file 1 | |
| Biological sample (Homo-sapiens) | Primary human peripheral blood mononuclear cells | This paper | Isolated from the patient and healthy controls |
|
| Antibody | Anti-human Phospho JNK (mouse monoclonal) |
Cell Signaling Technologies | Cat# 9,255 | Flow cytometry (1:100) |
| Antibody | Anti-Human STIM1(rabbit) | Cell Signaling Technologies | Cat# 5,668 S | WB (1:5000) |
| Antibody | Anti-His-tag(rabbit) | Cell Signaling Technologies | Cat# 12,698 S | WB (1:5000) |
| Antibody | Anti-FLAG tag(mouse monoclonal) |
Millipore Sigma | Cat# F3040 | WB (1:5000) |
| Antibody | Anti-human ORAI1(rabbit polyclonal) |
Millipore Sigma | Cat# AB9868 | WB (1:5000) |
| Antibody | Anti-Vav1(rabbit) | Cell Signaling Technologies | Cat# 2,502 | WB (1:2000) |
| Antibody | Anti-β-actin(mouse monoclonal) |
Santa Cruz Biotechnology | Cat# sc-47778 | WB (1:2500) |
| Antibody | anti-human CD3 antibody (mouse monoclonal) | Bio X Cell | Clone OKT-3 | 1 μg/ml |
| Antibody | Anti-human CD28 antibody (mouse monoclonal) | Bio X Cell | Cat. #: BE0921Clone CD28.2 | 1 μg/ml |
| Antibody | Anti-CD4-FITC(mouse monoclonal) |
eBioscience | Clone OKT-4 | FACS5 μl/test |
| Antibody | Anti-IFN- gama-PE-Cy7 |
eBioscience | Clone 45.B3 | FACS5 μl/test |
| Antibody | Ant-IL2-PE | eBioscience | Cat# MQ1-17H12 | FACS5 μl/test |
| Antibody | Anti-human,CD19-BV421(mouse monoclonal) |
BD | Cat. #: 562,440Clone HIB19 | FACS5 μl/test |
| Antibody | Anti-human, CD27-PE (mouse monoclonal) |
BD | Cat. #: No:555,441Clone: M-T271 | FACS20 μl/test |
| Antibody | Anti-human,IgD-FITC(mouse monoclonal) |
BD | Cat. #: 561,490Clone IA6-2 | FACS5 μl/test |
| Antibody | Anti-human,IgM-Perc-cy5.5(mouse monoclonal) |
BD | Cat. #: 561,285Clone G20-R7 | FACS5 μl/test |
| Antibody | Anti-human, CD24-FITC (mouse monoclonal) | BD | Cat. #: 555,427Clone ML-5 | FACS20 μl/test |
| Antibody | Anti-human,CD38-Pecy7(mouse monoclonal) |
BD | Cat. #: 335,825CloneHB-7 | FACS5 μl/test |
| Antibody | Anti-human,CD3-V500(mouse monoclonal) |
BD | Cat. #: 561,416Clone UCHT-1 | FACS5 μl/test |
| Antibody | Anti-human, CD4-BV421(mouse monoclonal) |
BD | Cat. #: 562,424Clone RPA-T4 | FACS5 μl/test |
| Antibody | Anti-human, CD25-Pecy7monoclonal |
BD | Cat. #: 335,824Clone 2A3 | FACS5 μl/test |
| Antibody | Anti-human, CD127-PrcP Cy5.5(mouse monoclonal) |
BD | Cat. #: 560,551Clone HIL-7R-M21 | FACS5 μl/test |
| Antibody | Anti-human, FoxP3- Alexa 488(mouse monoclonal) |
BD | Cat. #: 566,526Clone 2632/E7 | FACS5 μl/test |
| Antibody | Anti-human, CD196-PE(mouse monoclonal) |
BD | Cat. #: 559 562Clone 11A9 | FACS10 μl/test |
| Antibody | Anti-human,CD-183-APC(mouse monoclonal) |
BD | Cat. #: 550,967 | FACS20 μl/test |
| Antibody | Polyclonal F(ab′)two goat anti-human IgM/IgG/IgA |
Jackson ImmunoResearch | Cat. #: 109-006-129 | 10 μg/ml |
| Antibody | Anti-human CD138-APC(mouse monoclonal) |
Miltenyi Biotech | Cat. #: 130-117-395 | FACS2 μl/test |
| Recombinant DNA reagent |
FGllF | This paper | N/A | |
| Recombinant DNA reagent |
pmCherry-N1 | Clontech | Clontech plasmid#632,523 | |
| Recombinant DNA reagent |
pMD2.G | Addgene | Addgene plasmid #12,259 | |
| Recombinant DNA reagent |
psPAX2 | Addgene | Addgene plasmid #12,260 | |
| Recombinant DNA reagent |
pLentiCas9- blasticidin |
Addgene | Addgene plasmid #52,962 | |
| Recombinant DNA reagent |
pLentiguide-puro_ hCRACR2Asg#1 |
This paper | Details in Supplementary file 1 |
|
| Recombinant DNA reagent |
pLentiguide-puro_ hCRACR2Asg#2 |
This paper | Details in Supplementary file 1 |
|
| Recombinant DNA reagent |
pLentiguide-puro_ hCRACR2Asg#3 |
This paper | Details in Supplementary file 1 |
|
| Recombinant DNA reagent |
pLentiguide-puro_ hORAI1sg |
This paper | Details in Supplementary file 1 |
|
| Recombinant DNA reagent |
FG11F CRACR2A WT |
This paper | Details in Supplementary file 1 |
|
| Recombinant DNA reagent |
FG11F CRACR2AE278D | This paper | Details in Supplementary file 1 |
|
| Recombinant DNA reagent |
FG11F CRACR2AR144G | This paper | Details in Supplementary file 1 |
|
| Recombinant DNA reagent |
FG11F CRAC R2AR144G,E300* |
This paper | Details in Supplementary file 1 |
|
| Sequence- based reagent |
SureSelect XT Human All Exon V5 |
Agilent | ||
| Sequence- based reagent |
CRACR2AExon 6_F | This paper | ATGATTCCT GGCAGGTGAGA |
|
| Sequence- based reagent |
CRACR2AExon 6_R | This paper | ATTCCAGTG CAGGGACCAG |
|
| Sequence- based reagent |
CRACR2AExon 9_F | This paper | GGCCCTGATG TTGAGTAGGT |
|
| Sequence- based reagent |
CRACR2AExon 9_R | This paper | GTGAATGGC AGGGAAAGTGG |
|
| Sequence- based reagent |
CRACR2AExon 10_F | This paper | AAACAAGGT GAGGCCAGGG |
|
| Sequence- based reagent |
CRACR2AExon 10_R | This paper | AGCCCAAAT CCTCTT TTCACAG |
|
| Peptide, recombinant protein |
IL-2 | Roche | Cat. #: HIL2-RO | |
| Peptide, recombinant protein |
IL-21 | Peprotech | Cat. #: 200–21 | 50 ng/ml |
| Peptide, recombinant protein |
IL-2 | Peprotech | Cat# 200–02 | 20 units/ml |
| Commercial assay or kit |
aCD3 plates | Corning | Cat. #:354,725 | 10 ng/ml |
| Commercial assay or kit |
B cell isolation kit | Miltenyi Biotech, | Cat. #:130-091-151 | 100 U/ml |
| Commercial assay or kit |
QIAamp DNA Blood kit |
Qiagen | Cat. #:61,104 | |
| Commercial assay or kit |
Human IgG ELISA Quantitation Set | Bethyl Laboratories Inc, | Cat. #: E80-104 | |
| Commercial assay or kit |
Human IgM ELISA Quantitation Set | Bethyl Laboratories Inc, | Cat. #: E80-100 | |
| Commercial assay or kit |
MagniSort human naïve CD4+ T cell enrichment kit | Thermofisher Scientific | Cat. #: 8804-6814-74 | |
| Commercial assay or kit |
FOXP3/ Transcription Factor staining Buffer set |
Thermofisher Scientific | Cat. #: 2229155 | |
| Chemical compound, drug |
PHA-L | Sigma | Cat. #:L-4144 | |
| Chemical compound, drug |
3HThymidine | Perkin Elmer | Cat. #: 027001 | |
| Chemical compound, drug |
LymphoPrep | Axis Shield | Cat. #:1114547 | |
| Chemical compound, drug |
7-AAD-PerCP-Cy5 | BD | Cat. #: 559,925 | FACS5 μl/test |
| Chemical compound, drug |
Fura 2-AM | Thermofisher Scientific | Cat# F1221 | |
| Chemical compound, drug |
Brefeldin A | Thermofisher Scientific | Cat# 00-4506-51 | |
| Chemical compound, drug |
Thapsigargin | EMD Millipore | Cat# 80055–474 | |
| Chemical compound, drug |
Phorbol 12- myristate 13- acetate (PMA) |
EMD Millipore | Cat# 5.00582.0001 | |
| Chemical compound, drug |
Ionomycin | EMD Millipore | Cat# 407,951 | |
| Chemical compound, drug |
Polybrene | Millipore Sigma | Cat# TR-1003 | |
| Chemical compound, drug |
Puromycin | Invivogen | Cat# ant-pr-1 | |
| Chemical compound, drug |
Blasticidin | Invivogen | Cat# ant-bl-05 | |
| Chemical compound, drug |
Poly-D-Lysine | Thermo Fisher Scientific | Cat# A003E | |
| Chemical compound, drug |
Fixable Viability Dye eFluor 780 |
eBioscience | Cat# 65-0865-14 | 1 μl/ml of cells |
| Software, algorithm |
FlowJo v10 | TreeStar | ||
| Software, algorithm |
Slidebook software |
Intelligent Imaging Innovations, Inc | ||
| Software, algorithm |
OriginPro | Originlab | ||
| Software, algorithm |
Image J | NIH | ||
| Software, algorithm |
Fluoview FV10i Confocal Microscope | Olympus | ||
| Software, algorithm |
Fluoview software | FlowJo, LLC | ||
| Software, algorithm |
Exome sequence analysis (various) | Please see methods section | ||
| Other | LAS-3000 LCD camera | FujiFilm | ||
| Other | ECM 830 electroporator | BTX | ||
| Other | BD Fortessa flow cytometer | BD Biosciences | ||
| Other | Cytoflex LX flow cytometer | Beckman Coulter | ||
| Other | HiSeq 3,000 | Illumina |