REAGENT or RESOURCE	SOURCE	IDENTIFIER
Chemicals, peptides, and recombinant proteins
Q5 High-Fidelity Polymerase	NEB	Cat# M0491S
PolyJet	SignaGen	Cat# SL100688
DNase I	Thermo Fisher	Cat# EN0525
Poly-D-Lysine	Sigma-Aldrich	Cat# P6407
Lipofectamine LTX	Invitrogen	Cat# 15338500
Puromycin	Sigma-Aldrich	Cat# P8833
BSA	Roche	Cat# 10738328103
HEPES	Sigma-Aldrich	Cat# H3375
EDTA	Sigma-Aldrich	Cat# E6758
DAPI	Thermo Fisher	Cat# D21490
Forskolin	Calbiochem	Cat# 344281
IBMX	Sigma-Aldrich	Cat# I7018
Critical commercial assays
TOPO PCR Vector	Invitrogen	Cat# K450002
DNeasy Blood and Tissue Kit	QIAGEN	Cat# 69504
Q5 High-Fidelity PCR Kit	NEB	Cat #E0555S
PureLink Quick Gel Extraction Kit	Thermo Fisher	Cat# K220001
35-mm Glass bottom imaging dish	Cellvis	Cat# D35-20-1.5-N
Deposited data
FluoSTEP-ICUE sequence	GenBank	MT800777.1
FluoSTEP-AKAR sequence	GenBank	MT800778.1
Experimental models: Cell lines
HEK293T	ATCC	ATTC Cat# CRL-11268
HEK293A	Thermo Fisher	Cat# R70507
293-RIα	(Zhang et al., 2020)	N/A
Oligonucleotides
293-RIα cell line validation F	5′-TTTGTTGAAGTGGGAAGATTGG-3′	N/A
293-RIα cell line validation R	5′-TCAATAGGTGCTGGGATCTGC-3′	N/A
IDT Ultramer for generation of 293-RIα cell line	5′-GTTCTTGGCCCATGCTCAGACATCCTC AAACGAAACATACAGCAGTACAACAGTT TTGTGTCACTGTCTGTCGGTGGCGGCCG TGACCACATGGTCCTTCATGAGTATGTA AATGCTGCTGGGATTACATGAAATCTGC CTCCTGTGCCTCCCTTTTCTCCTCTCCCC AATCCATGCTTCACTCATGCAAACTGCTTTAT-3′	N/A
Recombinant DNA
px459	(Ran et al., 2013)	Addgene plasmid #62988
px459 human RIα gRNA	(Zhang et al., 2020)	N/A
pcDNA3.1 GFP1-10	(Kamiyama et al., 2016)	Addgene plasmid #70219
pcDNA3.1 FluoSTEP-AKAR	(Zhang et al., 2020)	N/A
pcDNA3.1 FluoSTEP-ICUE	(Zhang et al., 2020)	N/A
pcDNA3.1 FluoSTEP-AKAR(T/A)	(Zhang et al., 2020)	N/A
pcDNA3.1 FluoSTEP-ICUE(R279E)	(Zhang et al., 2020)	N/A
Software and algorithms
Prism	GraphPad	https://www.graphpad.com/scientific-software/prism/
ImageJ	NIH	https://imagej.nih.gov
Other
Zeiss Axiovert 200M Microscope	Carl Zeiss	N/A
ORCA-flash4.0LT digital CMOS camera	Hamamatsu	C11440-42U30
Lambda 10-2 filter changer	Sutter Instruments	LB10-B/IQ