Lentiviral vector integration in the human genome induces alternative splicing and generates aberrant transcripts.
Retroviral vectors integrate in genes and regulatory elements and may cause transcriptional deregulation of gene expression in target cells. Integration into transcribed genes also has the potential to deregulate gene expression at the posttranscriptional level by interfering with splicing and polya...
| Publicado en: | Journal of Clinical Investigation Vol. 122; no. 5; pp. 1653 - 1667 |
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| Autores principales: | , , , , , , , , , , , , , , , , , , , |
| Formato: | research Journal Article |
| Publicado: |
American Society for Clinical Investigation
May2012
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=104443540&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 104443540 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00219738 0YX jtl: Journal of Clinical Investigation issn: 00219738 maglogo: N pubinfo: dt: May2012 vid: 122 iid: 5 pid: 11983 pub: American Society for Clinical Investigation place: Ann Arbor, Michigan artinfo: ui: 104443540 NLM22523069 2011752200 10.1172/JCI61852 NLM22523069 PMC3347495 104443540 ppf: 1653 ppct: 14 formats: tig: atl: Lentiviral vector integration in the human genome induces alternative splicing and generates aberrant transcripts. aug: au: Moiani A Paleari Y Sartori D Mezzadra R Miccio A Cattoglio C Cocchiarella F Lidonnici MR Ferrari G Mavilio F Moiani, Arianna Paleari, Ylenia Sartori, Daniela Mezzadra, Riccardo Miccio, Annarita Cattoglio, Claudia Cocchiarella, Fabienne Lidonnici, Maria Rosa Ferrari, Giuliana Mavilio, Fulvio affil: Division of Genetics and Cell Biology, Istituto Scientifico H. San Raffaele, Milan, Italy sug: subj: RNA Genome, Human Retroviruses Physiology RNA Virus Infections Proteins Viral Physiology Nucleotides Cells Genes Retroviruses Documentation Cell Culture Techniques Biochemical Phenomena Polymerase Chain Reaction Funding Source Human ab: Retroviral vectors integrate in genes and regulatory elements and may cause transcriptional deregulation of gene expression in target cells. Integration into transcribed genes also has the potential to deregulate gene expression at the posttranscriptional level by interfering with splicing and polyadenylation of primary transcripts. To examine the impact of retroviral vector integration on transcript splicing, we transduced primary human cells or cultured cells with HIV-derived vectors carrying a reporter gene or a human β-globin gene under the control of a reduced-size locus-control region (LCR). Cells were randomly cloned and integration sites were determined in individual clones. We identified aberrantly spliced, chimeric transcripts in more than half of the targeted genes in all cell types. Chimeric transcripts were generated through the use of constitutive and cryptic splice sites in the HIV 5ι long terminal repeat and gag gene as well as in the β-globin gene and LCR. Compared with constitutively spliced transcripts, most aberrant transcripts accumulated at a low level, at least in part as a consequence of nonsense-mediated mRNA degradation. A limited set of cryptic splice sites caused the majority of aberrant splicing events, providing a strategy for recoding lentiviral vector backbones and transgenes to reduce their potential posttranscriptional genotoxicity. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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