Evaluation of the Elements of Short Hairpin RNAs in Developing shRNA-Containing CAR T Cells.
C-C chemokine receptor type 5 (CCR5)
Patients Living With HIV (PLWH)
Rev
Tat
chimeric antigen receptor (CAR)
hypoxanthine phosphoribosyltransferase 1 (HPRT)
shRNA-containing CARs
short hairpin RNA (shRNA)
Journal
Cancers
ISSN: 2072-6694
Titre abrégé: Cancers (Basel)
Pays: Switzerland
ID NLM: 101526829
Informations de publication
Date de publication:
20 May 2023
20 May 2023
Historique:
received:
25
04
2023
revised:
16
05
2023
accepted:
18
05
2023
medline:
22
6
2023
pubmed:
22
6
2023
entrez:
22
6
2023
Statut:
epublish
Résumé
Short hairpin RNAs (shRNAs) have emerged as a powerful tool for gene knockdown in various cellular systems, including chimeric antigen receptor (CAR) T cells. However, the elements of shRNAs that are crucial for their efficacy in developing shRNA-containing CAR T cells remain unclear. In this study, we evaluated the impact of different shRNA elements, including promoter strength, orientation, multiple shRNAs, self-targeting, and sense and antisense sequence composition on the knockdown efficiency of the target gene in CAR T cells. Our findings highlight the importance of considering multiple shRNAs and their orientation to achieve effective knockdown. Moreover, we demonstrate that using a strong promoter and avoiding self-targeting can enhance CAR T cell functionality. These results provide a framework for the rational design of CAR T cells with shRNA-mediated knockdown capabilities, which could improve the therapeutic efficacy of CAR T cell-based immunotherapy.
Identifiants
pubmed: 37345185
pii: cancers15102848
doi: 10.3390/cancers15102848
pmc: PMC10216594
pii:
doi:
Types de publication
Journal Article
Langues
eng
Références
Cell. 2018 Jul 26;174(3):549-563.e19
pubmed: 29937226
J Exp Med. 2018 Mar 5;215(3):985-997
pubmed: 29436394
N Engl J Med. 2018 Jul 5;379(1):64-73
pubmed: 29972754
Exp Hematol Oncol. 2023 Jan 27;12(1):14
pubmed: 36707873
Mol Ther. 2002 Oct;6(4):481-9
pubmed: 12377189
Mol Ther. 2008 Sep;16(9):1630-6
pubmed: 18665161
J Vis Exp. 2021 Mar 15;(169):
pubmed: 33779622
J Mol Biol. 1966 Aug;19(2):548-55
pubmed: 5969078
Methods Mol Biol. 2019;1937:235-258
pubmed: 30706401
Biomed Pharmacother. 2021 May;137:111339
pubmed: 33550044
Mol Ther Methods Clin Dev. 2020 Sep 28;19:285-294
pubmed: 33102620
Blood. 2005 Aug 1;106(3):818-26
pubmed: 15831707
Genomics Proteomics Bioinformatics. 2010 Sep;8(3):170-9
pubmed: 20970745
J Immunother Cancer. 2020 Apr;8(1):
pubmed: 32303620
Oncotarget. 2017 Jun 27;8(26):43048-43060
pubmed: 28574837
Stem Cell Res Ther. 2021 Jul 28;12(1):428
pubmed: 34321099
N Engl J Med. 2022 Feb 17;386(7):692-696
pubmed: 34904797
Mol Ther Methods Clin Dev. 2022 Apr 13;25:344-359
pubmed: 35573050
RNA. 2010 Jul;16(7):1328-39
pubmed: 20498457
JCI Insight. 2018 May 17;3(10):
pubmed: 29769444
Microbes Infect. 2021 May-Jun;23(4-5):104805
pubmed: 33711449
N Engl J Med. 2021 Feb 25;384(8):705-716
pubmed: 33626253
Brief Funct Genomics. 2020 May 20;19(3):175-182
pubmed: 31950135
Front Immunol. 2022 Jan 20;12:816515
pubmed: 35126374
J Virol. 1986 Aug;59(2):284-91
pubmed: 3016298
Methods Mol Biol. 2010;629:141-58
pubmed: 20387148
Science. 2000 Apr 7;288(5463):55-6
pubmed: 10766634
Biotechnol Prog. 2021 Jan;37(1):e3066
pubmed: 32808434
IUBMB Life. 2002 Feb;53(2):107-14
pubmed: 12049192
Viruses. 2010 Feb;2(2):574-600
pubmed: 21994649
AIDS. 1996 Dec;10 Suppl 3:S13-20
pubmed: 8970707
AIDS Res Hum Retroviruses. 1993 Oct;9(10):1031-7
pubmed: 7506552
Exp Hematol Oncol. 2020 Jun 08;9:11
pubmed: 32523801
Front Immunol. 2020 Jun 17;11:1109
pubmed: 32625204
Sci Rep. 2017 Apr 7;7(1):737
pubmed: 28389661
Mol Ther. 2008 Feb;16(2):378-86
pubmed: 18071333
J Immunother Cancer. 2022 Jan;10(1):
pubmed: 35027426
Cell Mol Immunol. 2021 Sep;18(9):2188-2198
pubmed: 34381179
BMC Biotechnol. 2018 Jan 30;18(1):4
pubmed: 29378552
Mol Ther. 2022 Feb 2;30(2):579-592
pubmed: 34628052
Front Immunol. 2022 Jan 03;12:811364
pubmed: 35046962
Nucleic Acids Res. 2015 Nov 16;43(20):e134
pubmed: 26130722
J Transl Med. 2022 May 23;20(1):239
pubmed: 35606821
J Immunother Cancer. 2017 Mar 21;5:26
pubmed: 28331616
Stem Cell Res Ther. 2021 Oct 7;12(1):527
pubmed: 34620233
Cancer Immunol Res. 2021 Jan;9(1):75-88
pubmed: 33093217
Mol Ther. 2006 Oct;14(4):494-504
pubmed: 16844419
Oncotarget. 2017 Dec 19;9(2):2268-2278
pubmed: 29416770
Cell. 2019 Oct 31;179(4):880-894.e10
pubmed: 31668804