Construction of an rAAV Producer Cell Line through Synthetic Biology.
HEK293
adeno-associated virus
biomanufacturing
synthetic biology
Journal
ACS synthetic biology
ISSN: 2161-5063
Titre abrégé: ACS Synth Biol
Pays: United States
ID NLM: 101575075
Informations de publication
Date de publication:
21 10 2022
21 10 2022
Historique:
pubmed:
12
10
2022
medline:
25
10
2022
entrez:
11
10
2022
Statut:
ppublish
Résumé
Recombinant adeno-associated viruses (rAAV) are important gene delivery vehicles for gene therapy applications. Their production relies on plasmid transfection or virus infection of producer cells, which pose a challenge in process scale-up. Here, we describe a template for a transfection-free, helper virus-free rAAV producer cell line using a synthetic biology approach. Three modules were integrated into HEK293 cells including an rAAV genome and multiple inducible promoters controlling the expression of AAV Rep, Cap, and helper coding sequences. The synthetic cell line generated infectious rAAV vectors upon induction. Independent control over replication and packaging activities allowed for manipulation of the fraction of capsid particles containing viral genomes, affirming the feasibility of tuning gene expression profiles in a synthetic cell line for enhancing the quality of the viral vector produced. The synthetic biology approach for rAAV production presented in this study can be exploited for scalable biomanufacturing.
Identifiants
pubmed: 36219557
doi: 10.1021/acssynbio.2c00207
pmc: PMC9595119
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3285-3295Références
J Virol. 1998 Mar;72(3):2224-32
pubmed: 9499080
J Virol. 1996 Sep;70(9):6497-501
pubmed: 8709289
Mol Ther. 2016 Feb;24(2):287-297
pubmed: 26437810
J Gene Med. 2000 Jul-Aug;2(4):260-8
pubmed: 10953917
J Virol. 1996 Mar;70(3):1845-54
pubmed: 8627709
Gene Ther. 1999 Jul;6(7):1322-30
pubmed: 10455443
Virol J. 2005 May 06;2:43
pubmed: 15877812
Hum Gene Ther. 2014 Mar;25(3):212-22
pubmed: 24299301
Hum Gene Ther. 2002 Nov 1;13(16):1935-43
pubmed: 12427305
Mol Ther Methods Clin Dev. 2019 Feb 16;13:279-289
pubmed: 30886878
Mol Ther. 2005 Dec;12(6):1217-25
pubmed: 16213797
Biotechnol J. 2021 Jan;16(1):e2000025
pubmed: 32975881
J Virol. 1995 Nov;69(11):6880-5
pubmed: 7474103
J Gen Virol. 1997 Sep;78 ( Pt 9):2131-8
pubmed: 9291998
J Virol. 2002 Feb;76(4):1904-13
pubmed: 11799185
Viruses. 2010 Aug;2(8):1681-1703
pubmed: 21994701
J Virol. 1998 Jan;72(1):420-7
pubmed: 9420241
J Virol. 2016 Jan 13;90(7):3411-27
pubmed: 26764008
Virus Res. 2016 Jan 2;212:39-52
pubmed: 26116898
J Virol. 2000 Oct;74(20):9441-50
pubmed: 11000213
Gene Ther. 1998 Jul;5(7):938-45
pubmed: 9813665
Annu Rev Virol. 2014 Nov;1(1):427-51
pubmed: 26958729
J Virol. 1994 Aug;68(8):4847-56
pubmed: 8035483
Proc Natl Acad Sci U S A. 2005 Sep 20;102(38):13634-9
pubmed: 16157891
Hum Gene Ther. 1995 Oct;6(10):1329-41
pubmed: 8590738
J Mol Med (Berl). 2021 May;99(5):593-617
pubmed: 33594520
Mol Ther Methods Clin Dev. 2018 Nov 01;11:180-190
pubmed: 30533449
Blood. 2013 Jul 4;122(1):23-36
pubmed: 23596044
Virology. 1989 Nov;173(1):120-8
pubmed: 2554565
Proc Natl Acad Sci U S A. 2010 Jun 1;107(22):10220-5
pubmed: 20479244
J Virol. 2000 Dec;74(24):11511-21
pubmed: 11090148
Mol Ther. 2010 Jan;18(1):80-6
pubmed: 19904234
Hum Gene Ther. 2009 Aug;20(8):796-806
pubmed: 19569968
Cell. 2006 Sep 8;126(5):995-1004
pubmed: 16959577
Science. 2019 Nov 29;366(6469):1139-1143
pubmed: 31780559
Mol Ther. 2000 Apr;1(4):376-82
pubmed: 10933956
J Virol. 2015 Nov 11;90(3):1278-89
pubmed: 26559843
Bioinformatics. 2010 Apr 1;26(7):966-8
pubmed: 20147306
J Virol. 2002 Dec;76(24):13015-27
pubmed: 12438627
Hum Gene Ther. 2009 Aug;20(8):861-70
pubmed: 19419276
J Pharm Sci. 2021 Jul;110(7):2609-2624
pubmed: 33812887
Cytotechnology. 1994;14(3):183-90
pubmed: 7765589
Nat Med. 2006 Mar;12(3):342-7
pubmed: 16474400
EMBO J. 2001 Jun 15;20(12):3282-91
pubmed: 11406604
Gene Ther. 2008 Jun;15(11):840-8
pubmed: 18418418
Mol Ther Methods Clin Dev. 2016 Mar 16;3:16002
pubmed: 27014711
J Virol. 1994 Nov;68(11):7169-77
pubmed: 7933099
J Gene Med. 2004 Dec;6(12):1369-81
pubmed: 15538729