دورية أكاديمية
أعرض في EDS

Rescue of splicing-mediated intron loss maximizes expression in lentiviral vectors containing the human ubiquitin C promoter.

التفاصيل البيبلوغرافية
العنوان: Rescue of splicing-mediated intron loss maximizes expression in lentiviral vectors containing the human ubiquitin C promoter.
المؤلفون: Cooper AR; Molecular Biology Interdepartmental PhD Program, University of California, Los Angeles, Los Angeles, CA 90095, USA., Lill GR; Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA., Gschweng EH; Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA., Kohn DB; Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA Department of Pediatrics, University of California, Los Angeles, Los Angeles, CA 90095, USA Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA dkohn1@mednet.ucla.edu.
المصدر: Nucleic acids research [Nucleic Acids Res] 2015 Jan; Vol. 43 (1), pp. 682-90. Date of Electronic Publication: 2014 Dec 17.
نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE
أسماء مطبوعة: Publication: 1992- : Oxford : Oxford University Press
Original Publication: London, Information Retrieval ltd.
مواضيع طبية MeSH: Genetic Vectors* , Introns* , Promoter Regions, Genetic*, Lentivirus/*genetics , Ubiquitin C/*genetics, Enhancer Elements, Genetic ; Exons ; Gene Expression ; HEK293 Cells ; Humans ; K562 Cells ; Peptide Elongation Factor 1/genetics ; RNA Splicing
مستخلص: Lentiviral vectors almost universally use heterologous internal promoters to express transgenes. One of the most commonly used promoter fragments is a 1.2-kb sequence from the human ubiquitin C (UBC) gene, encompassing the promoter, some enhancers, first exon, first intron and a small part of the second exon of UBC. Because splicing can occur after transcription of the vector genome during vector production, we investigated whether the intron within the UBC promoter fragment is faithfully transmitted to target cells. Genetic analysis revealed that more than 80% of proviral forms lack the intron of the UBC promoter. The human elongation factor 1 alpha (EEF1A1) promoter fragment intron was not lost during lentiviral packaging, and this difference between the UBC and EEF1A1 promoter introns was conferred by promoter exonic sequences. UBC promoter intron loss caused a 4-fold reduction in transgene expression. Movement of the expression cassette to the opposite strand prevented intron loss and restored full expression. This increase in expression was mostly due to non-classical enhancer activity within the intron, and movement of putative intronic enhancer sequences to multiple promoter-proximal sites actually repressed expression. Reversal of the UBC promoter also prevented intron loss and restored full expression in bidirectional lentiviral vectors.
(© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.)
References: Nature. 2010 Sep 16;467(7313):318-22. (PMID: 20844535)
RNA. 2013 Jun;19(6):811-27. (PMID: 23616639)
Mol Ther. 2006 Feb;13(2):391-400. (PMID: 16226060)
Gene. 2009 Dec 1;448(1):88-101. (PMID: 19733223)
J Biomed Sci. 2004 May-Jun;11(3):278-94. (PMID: 15067211)
Curr Opin Biotechnol. 2002 Oct;13(5):429-36. (PMID: 12459333)
J Virol. 2002 Jul;76(14):7209-19. (PMID: 12072520)
Annu Rev Biochem. 1998;67:509-44. (PMID: 9759496)
Mol Ther. 2013 Sep;21(9):1705-17. (PMID: 23857176)
Biochem Biophys Res Commun. 2007 Jun 29;358(2):521-7. (PMID: 17493584)
Gene Ther. 2010 Mar;17(3):400-11. (PMID: 19847204)
Science. 2002 Feb 1;295(5556):868-72. (PMID: 11786607)
Mol Ther. 2014 Mar;22(3):607-22. (PMID: 24256635)
Nat Biotechnol. 2005 Jan;23(1):108-16. (PMID: 15619618)
J Virol. 1998 Dec;72(12):9873-80. (PMID: 9811723)
J Clin Invest. 2008 Sep;118(9):3132-42. (PMID: 18688285)
PLoS One. 2010;5(7):e11834. (PMID: 20676373)
Science. 2013 Aug 23;341(6148):1233158. (PMID: 23845948)
J Clin Invest. 2013 Jul 1;:null. (PMID: 23863630)
J Virol. 1998 Nov;72(11):8463-71. (PMID: 9765382)
Mol Ther. 2000 Nov;2(5):458-69. (PMID: 11082319)
PLoS One. 2013;8(6):e65932. (PMID: 23776572)
Nat Biotechnol. 1997 Sep;15(9):871-5. (PMID: 9306402)
Mol Ther. 2002 Mar;5(3):242-51. (PMID: 11863413)
Science. 2003 Oct 17;302(5644):415-9. (PMID: 14564000)
Science. 2013 Aug 23;341(6148):1233151. (PMID: 23845947)
J Clin Invest. 2008 Sep;118(9):3143-50. (PMID: 18688286)
Science. 2009 Nov 6;326(5954):818-23. (PMID: 19892975)
Nat Med. 2010 Feb;16(2):198-204. (PMID: 20098431)
معلومات مُعتمدة: T32 GM007185 United States GM NIGMS NIH HHS; 2P01 HL073104 United States HL NHLBI NIH HHS; 2P01HL073104 United States HL NHLBI NIH HHS; GM007185 United States GM NIGMS NIH HHS
المشرفين على المادة: 0 (EEF1A1 protein, human)
0 (Peptide Elongation Factor 1)
0 (Ubiquitin C)
تواريخ الأحداث: Date Created: 20141219 Date Completed: 20150320 Latest Revision: 20181113
رمز التحديث: 20240628
مُعرف محوري في PubMed: PMC4288199
DOI: 10.1093/nar/gku1312
PMID: 25520191
قاعدة البيانات: MEDLINE
الوصف
تدمد:1362-4962
DOI:10.1093/nar/gku1312