Baboon Envelope Pseudotyped "Nanoblades" Carrying Cas9/gRNA Complexes Allow Efficient Genome Editing in Human T, B, and CD34
B cell
CRISPR/Cas9
T cell
gene editing
gene therapy
hematopoietic stem cells
immunotherapy
nanoblade
Journal
Frontiers in genome editing
ISSN: 2673-3439
Titre abrégé: Front Genome Ed
Pays: Switzerland
ID NLM: 101775540
Informations de publication
Date de publication:
2021
2021
Historique:
received:
09
09
2020
accepted:
18
01
2021
entrez:
29
10
2021
pubmed:
30
10
2021
medline:
30
10
2021
Statut:
epublish
Résumé
Programmable nucleases have enabled rapid and accessible genome engineering in eukaryotic cells and living organisms. However, their delivery into human blood cells can be challenging. Here, we have utilized "nanoblades," a new technology that delivers a genomic cleaving agent into cells. These are modified murine leukemia virus (MLV) or HIV-derived virus-like particle (VLP), in which the viral structural protein Gag has been fused to Cas9. These VLPs are thus loaded with Cas9 protein complexed with the guide RNAs. Highly efficient gene editing was obtained in cell lines, IPS and primary mouse and human cells. Here, we showed that nanoblades were remarkably efficient for entry into human T, B, and hematopoietic stem and progenitor cells (HSPCs) thanks to their surface co-pseudotyping with baboon retroviral and VSV-G envelope glycoproteins. A brief incubation of human T and B cells with nanoblades incorporating two gRNAs resulted in 40 and 15% edited deletion in the Wiskott-Aldrich syndrome (WAS) gene locus, respectively. CD34
Identifiants
pubmed: 34713246
doi: 10.3389/fgeed.2021.604371
pmc: PMC8525375
doi:
Types de publication
Journal Article
Langues
eng
Pagination
604371Informations de copyright
Copyright © 2021 Gutierrez-Guerrero, Abrey Recalde, Mangeot, Costa, Bernadin, Périan, Fusil, Froment, Martinez-Turtos, Krug, Martin, Benabdellah, Ricci, Giovannozzi, Gijsbers, Ayuso, Cosset and Verhoeyen.
Déclaration de conflit d'intérêts
PM and ER are named as inventors on a patent relating to the Nanoblades technology [patent applicants: Institut National de la Sante et de la Recherche Medicale (INSERM), Centre National de la Recherche Scientifique (CNRS)], Ecole Normale Superieure de Lyon, Universite Claude Bernard Lyon 1, Villeurb-Anne Cedex; application number: WO 2017/068077 Al; patent status: published, 27th April 2017. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Références
J Immunol. 2019 May 1;202(9):2546-2557
pubmed: 30867238
J Immunol. 2015 Sep 1;195(5):2493-501
pubmed: 26232436
Elife. 2017 Sep 28;6:
pubmed: 28956530
Mol Ther. 2017 May 3;25(5):1142-1154
pubmed: 28377044
Mol Ther Nucleic Acids. 2018 Dec 7;13:256-274
pubmed: 30317165
Nat Commun. 2019 Nov 19;10(1):5222
pubmed: 31745080
PLoS Pathog. 2016 Jun 09;12(6):e1005641
pubmed: 27281338
Microbiol Spectr. 2014 Aug 15;2(4):1-17
pubmed: 25215264
Nature. 2016 Jan 28;529(7587):490-5
pubmed: 26735016
Nucleic Acids Res. 2014 Dec 16;42(22):e168
pubmed: 25300484
N Engl J Med. 2011 Aug 25;365(8):725-33
pubmed: 21830940
Nucleic Acids Res. 2020 Aug 20;48(14):8178-8187
pubmed: 32619241
Cell. 2015 Oct 22;163(3):759-71
pubmed: 26422227
Mol Ther. 2011 Sep;19(9):1656-66
pubmed: 21750535
Science. 2014 Jan 3;343(6166):84-87
pubmed: 24336571
Cell Rep. 2016 Oct 25;17(5):1453-1461
pubmed: 27783956
Mol Ther Nucleic Acids. 2018 Sep 7;12:89-104
pubmed: 30195800
Mol Ther. 2018 Feb 7;26(2):456-467
pubmed: 29273498
Curr Gene Ther. 2017;16(5):297-308
pubmed: 28067165
Nat Biotechnol. 2014 Apr;32(4):347-55
pubmed: 24584096
Cell Death Dis. 2020 Nov 12;11(11):973
pubmed: 33184267
Nat Rev Immunol. 2006 Oct;6(10):741-50
pubmed: 16977339
Haematologica. 2019 Nov;104(11):e497-e501
pubmed: 31004018
Nat Med. 2018 Aug;24(8):1216-1224
pubmed: 30082871
Nat Biotechnol. 2015 Dec;33(12):1256-1263
pubmed: 26551060
J Exp Clin Cancer Res. 2018 Jul 21;37(1):163
pubmed: 30031396
Sci Transl Med. 2017 Jan 11;9(372):
pubmed: 28077679
Nat Commun. 2019 Oct 18;10(1):4730
pubmed: 31628330
Trends Biotechnol. 2013 Jul;31(7):397-405
pubmed: 23664777
Trends Biotechnol. 2015 Feb;33(2):132-40
pubmed: 25595557
Stem Cells. 2012 Aug;30(8):1771-80
pubmed: 22689616
Nature. 2016 Nov 17;539(7629):384-389
pubmed: 27820943
Nat Protoc. 2018 Feb;13(2):358-376
pubmed: 29370156
Sci Transl Med. 2011 Aug 24;3(97):97ra79
pubmed: 21865537
Blood. 2014 Feb 27;123(9):1422-4
pubmed: 24578496
Gene Ther. 2010 Apr;17(4):503-10
pubmed: 19956269
Front Immunol. 2019 Dec 16;10:2873
pubmed: 31921138
Virologie (Montrouge). 2019 Feb 1;23(1):3-6
pubmed: 31131829
Blood. 2009 Feb 12;113(7):1422-31
pubmed: 19059876
EMBO Mol Med. 2017 Nov;9(11):1574-1588
pubmed: 28899930
Cell Stem Cell. 2017 Oct 5;21(4):547-555.e8
pubmed: 28985529
Sci Rep. 2016 Feb 26;6:22167
pubmed: 26916217
Sci Transl Med. 2015 Mar 25;7(280):280ps7
pubmed: 25810311
Blood. 2002 Apr 1;99(7):2342-50
pubmed: 11895766
Curr Opin Mol Ther. 2006 Oct;8(5):390-5
pubmed: 17078381
Blood. 2014 Aug 21;124(8):1221-31
pubmed: 24951430
Nat Biotechnol. 2014 Sep;32(9):941-6
pubmed: 24952903
J Clin Invest. 2008 Sep;118(9):3143-50
pubmed: 18688286
J Thromb Haemost. 2016 Dec;14(12):2478-2492
pubmed: 27685947
Blood. 2017 Sep 28;130(13):1535-1542
pubmed: 28801449
Blood. 2018 Apr 26;131(17):1960-1973
pubmed: 29519807
Nature. 2015 Nov 12;527(7577):192-7
pubmed: 26375006
Cell Death Dis. 2015 Jul 23;6:e1831
pubmed: 26203860
Sci Immunol. 2019 May 17;4(35):
pubmed: 31101673
Mol Cell Pediatr. 2018 Nov 14;5(1):9
pubmed: 30430274
Nat Protoc. 2019 Jan;14(1):1-27
pubmed: 30559373
J Allergy Clin Immunol. 2020 Aug;146(2):262-265
pubmed: 32623069
Blood. 2012 Nov 22;120(22):4334-42
pubmed: 22898597
Blood Adv. 2019 Feb 12;3(3):461-475
pubmed: 30755435
Nat Commun. 2020 Jul 29;11(1):3778
pubmed: 32728076
Mol Ther. 2015 Jan;23(1):63-70
pubmed: 25189742
Cell Rep. 2018 May 29;23(9):2606-2616
pubmed: 29847792
Blood. 2006 Apr 15;107(8):3084-90
pubmed: 16397136
Nature. 2014 Jun 12;510(7504):235-240
pubmed: 24870228
Nat Genet. 2018 Apr;50(4):498-503
pubmed: 29610478
Cancers (Basel). 2020 Jul 15;12(7):
pubmed: 32679920
Bone Marrow Transplant. 2019 Dec;54(12):1940-1950
pubmed: 30903024
Nat Methods. 2010 Nov;7(11):929-35
pubmed: 20935652
Nat Commun. 2020 Mar 13;11(1):1334
pubmed: 32170079
Cytogenet Genome Res. 2002;98(2-3):126-35
pubmed: 12697994
Sci Rep. 2018 Aug 14;8(1):12144
pubmed: 30108345
Nat Commun. 2019 Jan 3;10(1):45
pubmed: 30604748
Nat Med. 2019 Sep;25(9):1396-1401
pubmed: 31501599
Int J Hematol. 2011 Apr;93(4):434-439
pubmed: 21384097
Blood. 2012 Mar 22;119(12):2709-20
pubmed: 22160384
Nat Commun. 2019 Apr 9;10(1):1634
pubmed: 30967552
Elife. 2019 Jan 17;8:
pubmed: 30648968
J Immunol Methods. 2018 Jun;457:33-40
pubmed: 29614266
Mol Ther. 2015 Nov;23(11):1734-1747
pubmed: 26281898
Nat Med. 2014 Oct;20(10):1101-3
pubmed: 25295939
Sci Transl Med. 2017 Oct 11;9(411):
pubmed: 29021165
J Invest Dermatol. 2020 Feb;140(2):338-347.e5
pubmed: 31437443
Cell Stem Cell. 2017 Nov 2;21(5):574-590
pubmed: 29100011
Science. 2012 Aug 17;337(6096):816-21
pubmed: 22745249
Bioessays. 2015 Nov;37(11):1202-14
pubmed: 26293289
Mol Ther. 2019 Jan 2;27(1):137-150
pubmed: 30424953
Nat Biotechnol. 2015 Sep;33(9):985-989
pubmed: 26121415
Cell. 2014 Oct 23;159(3):647-61
pubmed: 25307932
Science. 2014 Nov 28;346(6213):1258096
pubmed: 25430774
Clin Cancer Res. 2017 May 1;23(9):2255-2266
pubmed: 27815355
Nat Methods. 2018 Nov;15(11):941-946
pubmed: 30297964
Nat Med. 2019 May;25(5):776-783
pubmed: 30911135
Blood. 2009 Oct 1;114(14):2900-8
pubmed: 19643989
Hum Gene Ther. 2016 Oct;27(10):729-740
pubmed: 27483988
Genet Couns. 2016;27(1):9-24
pubmed: 27192888
J Clin Invest. 2014 Oct;124(10):4154-61
pubmed: 25271723
Mol Ther Methods Clin Dev. 2016 Mar 30;5:16019
pubmed: 27069952
Immunol Rev. 2010 Jul;236:125-38
pubmed: 20636813
J Immunol. 2018 Sep 1;201(5):1586-1598
pubmed: 30021769
Cell Stem Cell. 2019 Nov 7;25(5):607-621.e7
pubmed: 31543367
Mol Ther. 2011 Jan;19(1):28-35
pubmed: 21045810
Cell Stem Cell. 2014 Nov 6;15(5):643-52
pubmed: 25517468
Nat Commun. 2019 Sep 6;10(1):4045
pubmed: 31492863
Cell Stem Cell. 2019 Nov 7;25(5):592-593
pubmed: 31703769
Nat Methods. 2017 Jun;14(6):600-606
pubmed: 28459459
Blood Adv. 2017 Oct 24;1(23):2088-2104
pubmed: 29296856
PLoS One. 2013 Nov 11;8(11):e79047
pubmed: 24244415
J Exp Med. 2019 Jun 3;216(6):1301-1310
pubmed: 30975893
Sci Rep. 2018 Jan 17;8(1):888
pubmed: 29343825
EMBO Mol Med. 2018 Nov;10(11):
pubmed: 30224381
Blood. 2004 Mar 15;103(6):2079-87
pubmed: 14630817