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. 2021 Dec 14;3(1):101031. doi: 10.1016/j.xpro.2021.101031
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Armenian hamster anti-mouse TCR-β chain, APC/Fire750-conjugated BioLegend H57-597; Cat#109246; RRID: AB_2629697
Mouse anti-human CD45, ECD-conjugated Beckman Coulter J.33; Cat#A07784;
Mouse monoclonal anti-human CD8a, APC-eFluor® 780-conjugated Thermo Fisher Scientific OKT8; Cat#47-0086-42; RRID: AB_2573945
Mouse monoclonal anti-human CD8a, PE-conjugated Thermo Fisher Scientific OKT8; Cat#12-0086-42; RRID: AB_10732344
Mouse monoclonal anti-human TCR α/β, FITC-conjugated BioLegend IP26; Cat#306706; RRID: AB_314644
Purified monoclonal mouse anti-human CD28 BioLegend CD28.2; Cat#302902; RRID: AB_314304
Purified monoclonal mouse anti-human CD3 BioLegend OKT3; Cat#317302; RRID: AB_571927
Biological samples
Human blood/PMBCs Healthy volunteers Transfusionsmedizin, UK Erlangen
Chemicals, peptides, and recombinant proteins
Albumin from bovine serum (BSA) VWR Life Science Cat#A3294 CAS: 9048-46-8
Alt-R® Cas9 Electroporation Enhancer Integrated DNA Technologies Cat#1075916
Alt-R® S.p. Cas9 Nuclease V3 Integrated DNA Technologies Cat#1081059
ß-Mercaptoethanol (50 mM) Thermo Fisher Scientific Cat#31350010
DNA/RNA dye, peqGREEN VWR Peqlab Cat#732-3196
dNTP Mix (10 mM each) Thermo Fisher Scientific Cat#R01922
Dulbecco’s Phosphate Buffered Saline (PBS) Sigma-Aldrich Cat#D8537
GeneRuler 1 kb DNA Ladder Thermo Fisher Scientific Cat#SM0311
Gentamicin (50 mg/mL) Thermo Fisher Scientific Cat#15750060
Heat-Inactivated Fetal Bovine Serum (South America) Anprotec Cat#AC-SM-0027
HEPES Carl Roth Cat#HN77.3; CAS: 7365-45-9
Herculase II Fusion DNA Polymer Agilent Technologies Cat#600677
L-Glutamine Sigma-Aldrich Cat#G8540; CAS: 56-85-9
Penicillin-Streptomycin (10,000 U/mL) Thermo Fisher Scientific Cat#15140122;
Poly-L-glutamic acid sodium salt, mol wt 15,000–50,000 (PGA) Sigma-Aldrich Cat#P4761-100mg; CAS: 26247-79-0
Propidium Iodide Thermo Fisher Scientific Cat#P1304MP; CAS: 25535-16-4
Recombinant Human IL-15 PeproTech Cat#200-15; Accession Number: P40933
Recombinant Human IL-2 PeproTech Cat#200-02; Accession Number: P60568
Recombinant Human IL-7 PeproTech Cat#200-07; Accession Number: P13232
RPMI 1640 Medium Thermo Fisher Scientific Cat#21875034
RT-PCR Grade Water Thermo Fisher Scientific Cat#AM9935
Critical commercial assays
MinElute PCR Purification Kit Qiagen Cat#28004
P3 Primary Cell 4D-NucleofectorTM X Kit S Lonza Cat#V4XP-3032
P3 Primary Cell 96-well NucleofectorTM Kit (960 RCT) Lonza Cat#V4SP-3960
Oligonucleotides
Alt-R® CRISPR-Cas9 tracrRNA Integrated DNA Technologies Cat#1072534
hTRAC crRNA sequence (on antisense strand): 5′-AGAGTCT
CTCAGCTGGTACA-3′		 Ren et al. (2017) n/a
hTRAC HDR genomic forward primer:
5′-CTGCCTTTACTCTGCCAGAG-3′		 Müller et al. (2021) n/a
hTRAC HDR genomic reverse primer:
5′- CATCATTGACCAGAGCTCTG-3′		 Müller et al. (2021) n/a
hTRBC crRNA sequence:
5′-GGAGAATGACGAGTGGACCC-3′		 Schober et al. (2019) n/a
Recombinant DNA
HDR DNA template sequence Schober et al. (2019) n/a
Software and algorithms
FlowJo, Version 10.7.2 Becton Dickinson & Company (BD) https://www.flowjo.com/solutions/flowjo;
RRID: SCR_008520
GraphPad Prism 8.3.0 (538) GraphPad Software https://www.graphpad.com/; RRID: SCR_002798
Other
4D-NucleofectorTM Core Unit Lonza Cat#AAF-1002B
4D-NucleofectorTM X Unit Lonza Cat#AAF-1002X
BD LSRFortessa™ Cell Analyzer BD Biosciences n/a