Investigating CRISPR/Cas9 gene drive for production of disease-preventing prion gene alleles.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2022
2022
Historique:
received:
27
03
2022
accepted:
18
05
2022
entrez:
7
6
2022
pubmed:
8
6
2022
medline:
10
6
2022
Statut:
epublish
Résumé
Prion diseases are a group of fatal neurodegenerative disorders that includes chronic wasting disease, which affects cervids and is highly transmissible. Given that chronic wasting disease prevalence exceeds 30% in some endemic areas of North America, and that eventual transmission to other mammalian species, potentially including humans, cannot be ruled out, novel control strategies beyond population management via hunting and/or culling must be investigated. Prion diseases depend upon post-translational conversion of the cellular prion protein, encoded by the Prnp gene, into a disease-associated conformation; ablation of cellular prion protein expression, which is generally well-tolerated, eliminates prion disease susceptibility entirely. Inspired by demonstrations of gene drive in caged mosquito species, we aimed to test whether a CRISPR/Cas9-based gene drive mechanism could, in principle, promote the spread of a null Prnp allele among mammalian populations. First, we showed that transient co-expression of Cas9 and Prnp-directed guide RNAs in RK13 cells generates indels within the Prnp open-reading frame, indicating that repair of Cas9-induced double-strand breaks by non-homologous end-joining had taken place. Second, we integrated a ~1.2 kb donor DNA sequence into the Prnp open-reading frame in N2a cells by homology-directed repair following Cas9-induced cleavages and confirmed that integration occurred precisely in most cases. Third, we demonstrated that electroporation of Cas9/guide RNA ribonucleoprotein complexes into fertilised mouse oocytes resulted in pups with a variety of disruptions to the Prnp open reading frame, with a new coisogenic line of Prnp-null mice obtained as part of this work. However, a technical challenge in obtaining expression of Cas9 in the male germline prevented implementation of a complete gene drive mechanism in mice.
Identifiants
pubmed: 35671288
doi: 10.1371/journal.pone.0269342
pii: PONE-D-22-09021
pmc: PMC9173614
doi:
Substances chimiques
Prion Proteins
0
Prions
0
RNA, Guide
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0269342Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
J Biol Chem. 2005 Mar 25;280(12):11247-58
pubmed: 15618225
J Virol. 2006 Jan;80(2):596-604
pubmed: 16378962
Vet Res. 2019 Jan 22;50(1):6
pubmed: 30670087
J Vis Exp. 2017 Jun 16;(124):
pubmed: 28654078
Genesis. 2016 Jul;54(7):389-97
pubmed: 27124574
Nature. 2015 Jun 25;522(7557):478-81
pubmed: 26061765
EMBO J. 2007 Jan 24;26(2):538-47
pubmed: 17245436
Proc Natl Acad Sci U S A. 2015 Dec 8;112(49):E6736-43
pubmed: 26598698
FASEB J. 2020 Feb;34(2):2359-2375
pubmed: 31907995
Nat Methods. 2020 May;17(5):471-479
pubmed: 32203383
Cell. 1983 Nov;35(1):57-62
pubmed: 6414721
Science. 1982 Apr 9;216(4542):136-44
pubmed: 6801762
Nat Methods. 2013 Aug;10(8):741-3
pubmed: 23817069
Mol Reprod Dev. 2009 Jan;76(1):3
pubmed: 18951376
Vet Res. 2012 Dec 18;43:87
pubmed: 23249298
Transgenic Res. 2009 Apr;18(2):163-71
pubmed: 18821027
Nucleic Acids Res. 2020 Nov 4;48(19):10615-10631
pubmed: 32776089
Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):744-8
pubmed: 8570627
Nat Neurosci. 2010 Mar;13(3):310-8
pubmed: 20098419
Transgenic Res. 2015 Oct;24(5):921-7
pubmed: 26178246
Vaccine. 2018 Nov 29;36(50):7737-7743
pubmed: 30414779
Nat Biotechnol. 2016 Feb;34(2):137-8
pubmed: 26849513
J Biol Chem. 2019 Feb 22;294(8):2642-2650
pubmed: 30578300
Pathogens. 2018 Jan 01;7(1):
pubmed: 29301257
PLoS One. 2014 Dec 09;9(12):e114594
pubmed: 25490046
Genome Biol. 2017 Feb 20;18(1):35
pubmed: 28219395
Nature. 1992 Apr 16;356(6370):577-82
pubmed: 1373228
Nat Commun. 2020 Nov 3;11(1):5553
pubmed: 33144570
Nat Biotechnol. 2007 Jan;25(1):132-8
pubmed: 17195841
Front Mol Biosci. 2017 Apr 06;4:19
pubmed: 28428956
EMBO Mol Med. 2015 Feb 06;7(3):339-56
pubmed: 25661904
Mol Neurobiol. 1994 Apr-Jun;8(2-3):121-7
pubmed: 7999308
J Exp Med. 2016 Mar 7;213(3):313-27
pubmed: 26926995
Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10608-12
pubmed: 7902565
Cell Death Dis. 2015 Mar 19;6:e1697
pubmed: 25789972
BMC Vet Res. 2006 Jun 09;2:19
pubmed: 16764717
Nat Commun. 2017 Jan 09;8:13905
pubmed: 28067217
Genesis. 2018 May;56(5):e23212
pubmed: 29676032
Nat Biotechnol. 2016 Jan;34(1):78-83
pubmed: 26641531
Trends Cell Biol. 2016 Jan;26(1):52-64
pubmed: 26437586
PLoS One. 2011 Mar 28;6(3):e18047
pubmed: 21464935
PLoS One. 2016 Aug 30;11(8):e0161127
pubmed: 27575545
Nat Biotechnol. 2018 Dec;36(11):1062-1066
pubmed: 30247490
Arch Virol. 2008;153(9):1693-702
pubmed: 18696008
ACS Chem Biol. 2018 Feb 16;13(2):343-346
pubmed: 29400944
Emerg Infect Dis. 2017 Sep;23(9):1598-1600
pubmed: 28820384
Vaccine. 2015 Jan 29;33(5):726-33
pubmed: 25539804
J Cell Sci. 2008 Dec 15;121(Pt 24):4001-7
pubmed: 19056670
CRISPR J. 2021 Feb;4(1):19-24
pubmed: 33571044
J Clin Invest. 2014 Feb;124(2):847-58
pubmed: 24430187
Science. 2016 Jan 1;351(6268):84-8
pubmed: 26628643
J Mol Biol. 1999 Oct 1;292(4):797-817
pubmed: 10525406
Vet Med (Auckl). 2019 Oct 02;10:123-139
pubmed: 31632898
Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14945-9
pubmed: 8962161
Nature. 2019 Feb;566(7742):105-109
pubmed: 30675057
Cell. 1993 Jul 2;73(7):1339-47
pubmed: 8100741
JCI Insight. 2019 Jul 30;5:
pubmed: 31361599
Brain Pathol. 2019 Mar;29(2):248-262
pubmed: 30588682
Prion. 2017 Sep 3;11(5):368-380
pubmed: 28968152
BMC Genomics. 2008 Jul 28;9:352
pubmed: 18662396
J Biol Chem. 2018 Dec 21;293(51):19812-19822
pubmed: 30397182
Cell. 1998 Apr 17;93(2):203-14
pubmed: 9568713