In vivo and in vitro knockout system labelled using fluorescent protein via microhomology-mediated end joining.
Animals
CRISPR-Associated Protein 9
/ metabolism
CRISPR-Cas Systems
Carnitine O-Acetyltransferase
/ genetics
Cell Line, Tumor
Cell Survival
/ genetics
DNA End-Joining Repair
/ genetics
Dependovirus
/ genetics
Gene Editing
/ methods
Gene Knock-In Techniques
/ methods
Gene Knockout Techniques
/ methods
Genetic Vectors
Green Fluorescent Proteins
/ genetics
Kelch-Like ECH-Associated Protein 1
/ genetics
Male
Mice
Mice, Inbred C57BL
NIH 3T3 Cells
Retinal Ganglion Cells
/ metabolism
Staining and Labeling
/ methods
Staphylococcus aureus
/ enzymology
Transfection
Journal
Life science alliance
ISSN: 2575-1077
Titre abrégé: Life Sci Alliance
Pays: United States
ID NLM: 101728869
Informations de publication
Date de publication:
01 2020
01 2020
Historique:
received:
20
08
2019
revised:
12
12
2019
accepted:
12
12
2019
entrez:
26
12
2019
pubmed:
26
12
2019
medline:
12
1
2021
Statut:
epublish
Résumé
Gene knockout is important for understanding gene function and genetic disorders. The CRISPR/Cas9 system has great potential to achieve this purpose. However, we cannot distinguish visually whether a gene is knocked out and in how many cells it is knocked out among a population of cells. Here, we developed a new system that enables the labelling of knockout cells with fluorescent protein through microhomology-mediated end joining-based knock-in. Using a combination with recombinant adeno-associated virus, we delivered our system into the retina, where the expression of
Identifiants
pubmed: 31874862
pii: 3/1/e201900528
doi: 10.26508/lsa.201900528
pmc: PMC6932181
pii:
doi:
Substances chimiques
Keap1 protein, mouse
0
Kelch-Like ECH-Associated Protein 1
0
Green Fluorescent Proteins
147336-22-9
Carnitine O-Acetyltransferase
EC 2.3.1.7
CRISPR-Associated Protein 9
EC 3.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2019 Katayama et al.
Références
Arch Ophthalmol. 1981 Jan;99(1):137-43
pubmed: 7458737
Mol Ther Methods Clin Dev. 2017 Dec 01;8:87-104
pubmed: 29326962
Genome Biol. 2016 Jul 05;17(1):148
pubmed: 27380939
J Neurosci Res. 2012 Apr;90(4):802-15
pubmed: 22065590
BMC Genomics. 2016 Nov 28;17(1):979
pubmed: 27894274
Nature. 2015 Apr 9;520(7546):186-91
pubmed: 25830891
Proc Natl Acad Sci U S A. 2017 May 16;114(20):E3974-E3983
pubmed: 28465430
Mol Cell Biol. 2004 Aug;24(16):7130-9
pubmed: 15282312
Nature. 2015 Jan 29;517(7536):583-8
pubmed: 25494202
Invest Ophthalmol Vis Sci. 2014 Oct 02;55(11):7126-36
pubmed: 25277230
Prog Lipid Res. 2010 Jan;49(1):61-75
pubmed: 19720082
Development. 2017 May 1;144(9):1698-1711
pubmed: 28356311
Nature. 2012 Feb 15;482(7385):331-8
pubmed: 22337052
Hum Mol Genet. 2018 Apr 15;27(8):1486-1496
pubmed: 29452408
Angew Chem Int Ed Engl. 2016 May 23;55(22):6452-6
pubmed: 27079176
Nat Biotechnol. 2013 Sep;31(9):827-32
pubmed: 23873081
PLoS One. 2017 Dec 20;12(12):e0186678
pubmed: 29261660
Sci Rep. 2015 Mar 05;5:8841
pubmed: 25740433
Nephrol Dial Transplant. 2014 Apr;29(4):783-91
pubmed: 24523358
Annu Rev Biochem. 1988;57:261-83
pubmed: 3052273
J Cell Biol. 2005 Oct 24;171(2):313-25
pubmed: 16247030
Nat Commun. 2018 Mar 5;9(1):939
pubmed: 29507284
Sci Rep. 2017 Mar 14;7:44229
pubmed: 28290486
Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):1876-81
pubmed: 11854488
Vision Res. 2008 Feb;48(3):377-85
pubmed: 17950399
Nat Commun. 2014 Nov 20;5:5560
pubmed: 25410609
Nat Protoc. 2016 Jan;11(1):118-33
pubmed: 26678082
Mol Aspects Med. 2004 Oct-Dec;25(5-6):533-49
pubmed: 15363640
Genes Dev. 1999 Jan 1;13(1):76-86
pubmed: 9887101
ACS Synth Biol. 2016 May 20;5(5):395-404
pubmed: 26883397
Mol Neurobiol. 2016 Dec;53(10):6644-6652
pubmed: 26643543
Nature. 2010 Nov 4;468(7320):67-71
pubmed: 21048762
Science. 2012 Aug 17;337(6096):816-21
pubmed: 22745249
J Neurochem. 2013 Dec;127(5):669-80
pubmed: 23721546
Invest Ophthalmol Vis Sci. 2005 Sep;46(9):3188-96
pubmed: 16123418
Microbiology. 2009 Mar;155(Pt 3):733-40
pubmed: 19246744
Science. 2013 Feb 15;339(6121):823-6
pubmed: 23287722
Sci Rep. 2018 Aug 9;8(1):11930
pubmed: 30093719
Ann N Y Acad Sci. 2005 Aug;1053:153-61
pubmed: 16179519
Science. 2013 Feb 15;339(6121):819-23
pubmed: 23287718
Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):20038-43
pubmed: 19060199
Sci Rep. 2015 Dec 16;5:18141
pubmed: 26670005