Reagent/Resource	Reference or Source	Identifier or catalogue number
Experimental models
HEK‐293 Flp‐In cells (H. sapiens)	ThermoFisher	R75007
HEK‐293T cells (H. sapiens)	ATCC	CRL‐3216
Electrocompetent E. cloni cells (10G ELITE)	Lucigen	60052‐4
CFBE cells (H. sapiens)	Molinski et al (2015) Botelho et al (2015)	N/A
Recombinant DNA
MaMTH reporter vector using the pcDNA3.1(−) backbone	Gibson et al (2009) and Saraon et al (2020)	N/A
MaMTH bait destination vector	Gateway cloning technology (Thermo Fisher) and Saraon et al (2020)	N/A
wt‐ and F508del‐CFTR cDNA entry constructs	Dr. Johanna Rommens lab	Ref Seq: NM_000492.3
MaMTH prey destination vector	Saraon et al (2020)	N/A
Plasmid library generation	Human ORFeome V8.1 collection (Yang et al, 2011)	N/A
Antibodies
Anti‐Flag antibody	Sigma	F‐1804
Anti‐mouse IgG antibody conjugated with Alexa 647	Invitrogen	A‐31571
Anti‐CFTR 596	Cystic Fibrosis Foundation	N/A
Anti‐calnexin	BD Transduction Laboratories	610523
HRP‐conjugated goat anti‐mouse IgG	Bio‐Rad	170‐6516
Anti‐FLAG antibody	Millipore Sigma	F3165
Anti‐V5 antibody	Cell Signaling Technology	13202S
Oligonucleotides and other sequence‐based reagents
PCR primers	This study	Table EV1
Custom library of 421 Ambion Silencer Select siRNAs	This study	Table EV3
Non‐targeting Neg1 negative control siRNA	Ambion	4390844
Chemicals, enzymes and other reagents
DMEM	Wisent	319‐005‐CL
DMEM high glucose	Corning	10‐013‐CV
Opti‐MEM	Gibco	31985070
IntestiCultTM Human OGM	STEMCELL Technologies	06010
FBS	Gibco	10270
Hygromycin	Bioshop	HYG003.10
Blasticidin	Invivogen	ant‐bl
Puromycin	Invivogen	ant‐pr
Doxycycline	Sigma	9891
Primocin	Invivogen	ant‐pm
Vancomycin	Sigma	SBR00001
Gentamycin	Sigma	G1272
Y‐2632 (ROCK pathway inhibitor)	Sigma	SCM075
Forskolin	Sigma Aldrich	F3917
VX‐661	Selleckchem	S7059
VX‐809	Selleckchem	S1565
VX‐445	Selleckchem	S8851
CFTRinh‐172	Cystic Fibrosis Foundation EMD Millipore Corp	N/A
Amiloride	Spectrum Chemical	TCI‐A2599
Phire Tissue Direct Dilution Buffer	Thermo Scientific	N/A
Phire Tissue Direct PCR Master Mix	Thermo Scientific	N/A
QIAquick PCR Purification Kit	Qiagen	N/A
Qiagen Mini Kit	Qiagen	27106
Qiagen Micro Kit	Qiagen	74004
iSCRIPT cDNA synthesis kit	Bio‐Rad	1708890
SsofastTM EvaGreen fluorophore	Bio‐Rad	172‐5200
CellMask	Invitrogen	H32712
ER‐Tracker	Invitrogen	E34250
Hoechst 33342	Sigma	B2261
Calcein green	Invitrogen	C34852
X‐tremeGENE 9 DNA Transfection Reagent	Roche	6365787001
Lipofectamine 2000	Invitrogen	11668019
Lipofectamine LTX	Thermo Fisher Scientific	15338100
Laemmli sample buffer	Bio‐Rad	161‐0747
Protease inhibitor cocktail	Roche	11873580001
Clarity Western ECL Substrate	Bio‐Rad	1705061
Blue membrane potential dye	Molecular devices	R8034
384 well plates	Falcon	736‐2044
96 well plates	Sarstedt	83.3924
12 well plates	SPL Life Sciences	30012
PureCol®‐coated Transwell	Corning	CostarTM 3470
Non‐perfused Ussing chamber	Physiologic Instruments	N/A
Drying pearls	Sigma‐Aldrich	94098
μMACSTM FLAG magnetic microbeads and columns	Miltenyi Biotec	N/A
Matrigel	BD Biosciences	N/A
Software
GraphPad Prism	https://www.graphpad.com	N/A
Integrated Interactions Database (IID)	PMID: 34755877 http://ophid.utoronto.ca/iid	Version 2021‐05
NaViGaTOR	http://navigator/ophid.utoronto.ca/navigatorwp	Version 3.0.15
R programming language/Bowtie 2 Alignment Tool	R Core Team, Bowtie2 alignment tool (Langmead & Salzberg, 2012)	N/A
CellProfiler	McQuin et al (2018)	N/A
Image Lab	Bio‐Rad	N/A
MATLAB‐based organoid swelling capturing program	MATLAB	N/A
Other
NexteraXT library preparation kit	Illumina	N/A
NextSeq500 System	Illumina	N/A
HiSeq2500 System	Illumina	N/A
BD FACS Melody	BD Biosciences	N/A
Rank product method	Breitling et al (2004)	N/A
Genevac miVac Duo	Thermo Fisher Scientific	N/A
MultidropTM Combi peristaltic dispenser	Thermo Scientific	5840300
Leica DMI 6000B	Leica	N/A
ChemiDoc XRS	Bio‐Rad	N/A
Cellomics ArrayScan, VTI HCS Reader	Thermo Fisher Scientific	N/A
NanoDrop 2000	Thermo Fisher Scientific	N/A