Episodic live imaging of cone photoreceptor maturation in GNAT2-EGFP retinal organoids.
CRISPR
Cone photoreceptor
Fluorescent reporter
GNAT2
Live imaging
Retinal organoid
iPSC
Journal
Disease models & mechanisms
ISSN: 1754-8411
Titre abrégé: Dis Model Mech
Pays: England
ID NLM: 101483332
Informations de publication
Date de publication:
01 Nov 2023
01 Nov 2023
Historique:
received:
20
03
2023
accepted:
12
10
2023
medline:
22
11
2023
pubmed:
30
10
2023
entrez:
30
10
2023
Statut:
ppublish
Résumé
Fluorescent reporter pluripotent stem cell-derived retinal organoids are powerful tools to investigate cell type-specific development and disease phenotypes. When combined with live imaging, they enable direct and repeated observation of cell behaviors within a developing retinal tissue. Here, we generated a human cone photoreceptor reporter line by CRISPR/Cas9 genome editing of WTC11-mTagRFPT-LMNB1 human induced pluripotent stem cells (iPSCs) by inserting enhanced green fluorescent protein (EGFP) coding sequences and a 2A self-cleaving peptide at the N-terminus of guanine nucleotide-binding protein subunit alpha transducin 2 (GNAT2). In retinal organoids generated from these iPSCs, the GNAT2-EGFP alleles robustly and exclusively labeled immature and mature cones. Episodic confocal live imaging of hydrogel immobilized retinal organoids allowed tracking of the morphological maturation of individual cones for >18 weeks and revealed inner segment accumulation of mitochondria and growth at 12.2 μm3 per day from day 126 to day 153. Immobilized GNAT2-EGFP cone reporter organoids provide a valuable tool for investigating human cone development and disease.
Identifiants
pubmed: 37902188
pii: 335436
doi: 10.1242/dmm.050193
pmc: PMC10690052
pii:
doi:
Substances chimiques
enhanced green fluorescent protein
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NICHD NIH HHS
ID : T32 HD060549
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA137124
Pays : United States
Organisme : NEI NIH HHS
ID : R01 EY026661
Pays : United States
Organisme : NIH HHS
ID : 5R01CA137124
Pays : United States
Organisme : NIH HHS
ID : 5T32HD060549
Pays : United States
Organisme : NIH HHS
ID : R01EY026661
Pays : United States
Commentaires et corrections
Type : UpdateOf
Informations de copyright
© 2023. Published by The Company of Biologists Ltd.
Déclaration de conflit d'intérêts
Competing interests J.B. and D.C. are listed as inventors on a provisional patent application entitled ‘Fluorescent Cone Reporter iPSC Cells’ filed by Children's Hospital Los Angeles.
Références
Development. 1994 Aug;120(8):2091-102
pubmed: 7925013
Exp Eye Res. 2018 Jun;171:111-118
pubmed: 29518352
Invest Ophthalmol Vis Sci. 2017 Jul 1;58(9):3311-3318
pubmed: 28672397
Stem Cells. 2015 Dec;33(12):3504-18
pubmed: 26235913
Nature. 2014 Oct 16;514(7522):385-8
pubmed: 25252974
Stem Cell Reports. 2018 Sep 11;11(3):665-680
pubmed: 30100409
Prog Retin Eye Res. 2009 Jul;28(4):289-302
pubmed: 19501669
Biophys J. 2003 Feb;84(2 Pt 1):1317-27
pubmed: 12547812
Front Cell Dev Biol. 2022 Apr 28;10:870441
pubmed: 35573687
Transl Vis Sci Technol. 2020 Feb 18;9(3):21
pubmed: 32714647
Front Bioeng Biotechnol. 2018 Dec 18;6:199
pubmed: 30619846
J Comp Neurol. 2019 Jan 1;527(1):38-51
pubmed: 28074469
Stem Cells Transl Med. 2017 Nov;6(11):1972-1986
pubmed: 29024560
Invest Ophthalmol Vis Sci. 1997 Jan;38(1):196-206
pubmed: 9008644
Sci Rep. 2018 Feb 5;8(1):2370
pubmed: 29402929
Sci Rep. 2017 Apr 10;7(1):766
pubmed: 28396597
Development. 2019 Jan 9;146(1):
pubmed: 30567931
Proc Natl Acad Sci U S A. 2018 Oct 2;115(40):E9391-E9400
pubmed: 30213853
Dev Cell. 2017 Dec 18;43(6):763-779.e4
pubmed: 29233477
Ophthalmic Genet. 2023 Oct 9;:1-15
pubmed: 37807874
Int J Biol Macromol. 2016 May;86:917-28
pubmed: 26893053
Elife. 2019 Aug 27;8:
pubmed: 31451149
Prog Retin Eye Res. 2019 Mar;69:1-37
pubmed: 30445193
Sci Rep. 2020 Jun 24;10(1):10275
pubmed: 32581233
Mol Biol Cell. 2017 Oct 15;28(21):2854-2874
pubmed: 28814507
Nat Commun. 2015 Feb 19;6:6286
pubmed: 25695148
Nat Commun. 2014 Jun 10;5:4047
pubmed: 24915161
J Clin Invest. 2022 Jun 15;132(12):
pubmed: 35482419
Annu Rev Vis Sci. 2020 Sep 15;6:91-114
pubmed: 32936736
Front Cell Dev Biol. 2021 Nov 16;9:764725
pubmed: 34869356
Development. 2020 Dec 24;147(24):
pubmed: 33361444
Sci Rep. 2022 Apr 22;12(1):6646
pubmed: 35459774
Stem Cell Reports. 2017 Dec 12;9(6):1898-1915
pubmed: 29153988
Genomics. 1997 Aug 15;44(1):101-9
pubmed: 9286705
Nat Protoc. 2013 Nov;8(11):2281-2308
pubmed: 24157548
Genomics. 1993 Aug;17(2):442-8
pubmed: 8406495
Anal Biochem. 2007 Sep 1;368(1):95-9
pubmed: 17586454
Stem Cells. 2022 Mar 16;40(2):190-203
pubmed: 35293574
Dev Cell. 2020 May 18;53(4):473-491.e9
pubmed: 32386599