TRPA5 encodes a thermosensitive ankyrin ion channel receptor in a triatomine insect

. 2024 Mar 20;27(4):109541. doi: 10.1016/j.isci.2024.109541

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies

Monoclonal anti-FLAG M2 antibody produced in mouse	Sigma-Aldrich	Cat#F1804; RRID:AB_262044
Sheep Anti-Mouse IgG - Horseradish Peroxidase	Cytiva	Cat#NA931; RRID:AB_772210
Rabbit Anti-Sodium Potassium ATPase monoclonal antibody, unconjugated, clone EP1845Y	Abcam	Cat#ab76020; RRID:AB_1310695

Bacterial and virus strains

Stbl3 competent E. coli cells	ThermoFisher	Cat#C737303

Biological samples

Rhodnius prolixus antennae	Prof. Ian Orchard, University of Toronto Mississauga, Canada	RRID:NCBITaxon_13249
Rhodnius prolixus live adults	BEI	https://www.beiresources.org/Catalog/BEIVectors/NR-44077.aspx; RRID:NCBITaxon_13249

Chemicals, peptides, and recombinant proteins

Lipofectamine 2000	Life Technologies	Cat#11668019
Opti-MEM I Reduced Serum	Life Technologies	Cat#31985070
Accutase	Stemcell Technologies	Cat#07920
DMEM High Glucose, GlutaMAX	Life Technologies	Cat#10566016
Fetal Serum Bovine, Seradigm Premier	VWR	Cat#97068-085

Critical commercial assays

Direct-zol RNA kit	Zymo Research	Cat#R2051
GoScript™ Reverse Transcriptase kit	Promega	Cat#A5003
Advantage® 2 PCR Kit	Takara Bio	Cat#639206
Monarch® DNA gel extraction kit	NEB	Cat#T1020S
Qiaprep spin Miniprep	Qiagen	Cat#27104
endo-free ZymoPURE™ II Plasmid Midiprep Kit	Zymo Research	Cat#D4201
Pierce Cell surface Protein isolation kit	Thermo Scientific	Cat#89881
Trans-Blot Turbo RTA Mini 0.2 μm Nitrocellulose Transfer kit	Bio-Rad	Cat#1704270

Deposited data

Sequence Read Archive (SRA) PRJNA281760/SRA:SRP057515 at NCBI	Latorre-estivalis et al.⁴⁵	https://www.ncbi.nlm.nih.gov/sra/?term=SRP057515
Sequence Read Archive (SRA) PRJEB13049/SRA:ERP014587 at NCBI	NCBI	https://www.ncbi.nlm.nih.gov/sra/?term=ERP014587
Sequence Read Archive (SRA) PRJNA191820/SRA:SRP006783 at NCBI	NCBI	https://www.ncbi.nlm.nih.gov/sra/?term=SRP006783
R. prolixus genome assembly version RproC3.0.3	Mesquita et al.³⁶	VectorBase.org
dTRPA1 structure state 1	Wang et al.⁵⁹	PDB: 7YKR
dTRPA1 structure state 2	Wang et al.⁵⁹	PDB: 7YKS
hTRPA1 structure	Zhao et al.⁶¹	PDB: 6V9Y

Experimental models: Cell lines

Human: HEK293T	Sigma-Aldrich	Cat#12022001; RRID:CVCL_0063

Oligonucleotides

Primers for TRP amplification, subcloning, sanger verification and quantitative PCR analyses	See Table S7	N/A

Recombinant DNA

Plasmid DNA, rTRPV1 in pcDNA3	Prof. Rachelle Gaudet, MCB department, Harvard University	N/A
Plasmid DNA, dTRPA1-D in pcDNA3	Zhong et al.²²	N/A
Expression vector pcDNA5-FLAG-T2A-mRuby2	Lienard et al.⁵¹	N/A
Plasmid DNA, RpTRPA5B in pcDNA5-FLAG-T2A-mRuby	This paper	N/A
Plasmid DNA, rTRPV1 in pcDNA5-FLAG-T2A-mRuby2	This paper	N/A
Plasmid DNA, dTRPA1-D in pcDNA5-FLAG-T2A-mRuby2	This paper	N/A

Software and algorithms

NCBI-Blast	Altschul et al.¹⁰⁴	https://blast.ncbi.nlm.nih.gov/Blast.cgi
MAFFT	Katoh and Standley¹⁰⁷	https://mafft.cbrc.jp/alignment/server/index.html
IQ-Tree v1.6.11	Nguyen et al.¹⁰⁸ Trifinopoulos et al.¹⁰⁹ Kalyaanamoorthy et al.¹¹⁰	http://www.iqtree.org
R studio (R V3.6.3) ggtree package	Yu¹¹¹	https://bioconductor.org/packages/release/bioc/html/ggtree.html
Evolview	Zhang et al.¹¹²	https://evolgenius.info/evolview-v2/
Cutadapt version 1.16	Martin¹¹³	https://cutadapt.readthedocs.io/en/v1.16/
HISAT2 version 2.2.0	Kim et al.¹¹⁴	http://daehwankimlab.github.io/hisat2/download/
StringTie version 2.1.3b	Pertea et al.¹¹⁵	https://ccb.jhu.edu/software/stringtie/
Trinotate version 3.2.1	Bryant et al.¹¹⁶	https://github.com/Trinotate/Trinotate/releases
Apollo gene browser	Lee et al.¹¹⁷	http://genomearchitect.readthedocs.io
R studio (R V3.6.3) tximport package	Soneson et al.¹¹⁸	https://bioconductor.org/packages/release/bioc/html/tximport.html
Geneious	Kearse et al.¹¹⁹	https://www.geneious.com
CFX Maestro Software 2.3 (Biorad)	N/A	https://www.bio-rad.com/en-be/category/qpcr-analysis-software?ID=42a6560b-3ad7-43e9-bb8d-6027371de67a
AlphaFold2	Jumper et al.⁵⁶ Mirdita et al.⁵⁸	https://alphafold.ebi.ac.uk
PyMOL	Pymol¹²⁰	https://pymol.org/2/
DALI server	Holm¹²¹	http://ekhidna2.biocenter.helsinki.fi/dali/
Labview	Emerson	https://www.ni.com/en.html
Digidata 1440A ,pClamp10 software	Molecular Devices	https://info.molecularmolecul.com

Other

DOI link for source data related to Figure 1	This paper	https://figshare.com/s/2d1ad932f34bdca2a769
DOI link for source data related to Figure 2	This paper	https://figshare.com/s/995be46d32030cd52910
DOI link for source data related to Figures 3 and S7–S10	This paper	https://figshare.com/s/0606a76fe5e96202ef73
DOI link for source data and code related to Figure S1	This paper	https://figshare.com/s/aca16873b4cc7e62d242
DOI link for source data and code related to Figure S2	This paper	https://figshare.com/s/014ec6a509c3c3d01896
DOI link for source data and code related to Figure S3	This paper	https://figshare.com/s/09aa5dfd07dc56ee1bcc
DOI link for source data related to Figure S4	This paper	https://figshare.com/s/85aa2e7bfcd7a3bc4168
DOI link for source data related to Figure S5	This paper	https://figshare.com/s/995be46d32030cd52910
DOI link for source data related to Figure S6	This paper	https://figshare.com/s/789896f4ab3b1804a648