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Efficient character...
Efficient characterization of retro-, lenti-, and foamyvector-transduced cell populations by high-accuracy insertion site sequencing
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Schmidt, M (author)
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Glimm, H (author)
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Wissler, M (author)
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Hoffmann, G (author)
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- Olsson, Karin (author)
- Lund University,Lunds universitet,Avdelningen för molekylärmedicin och genterapi,Institutionen för laboratoriemedicin,Medicinska fakulteten,Division of Molecular Medicine and Gene Therapy,Department of Laboratory Medicine,Faculty of Medicine
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Sellers, S (author)
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Carbonaro, D (author)
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Tisdale, JF (author)
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Leurs, C (author)
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Hanenberg, H (author)
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Dunbar, CE (author)
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Kiem, HP (author)
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- Karlsson, Stefan (author)
- Lund University,Lunds universitet,Avdelningen för molekylärmedicin och genterapi,Institutionen för laboratoriemedicin,Medicinska fakulteten,Division of Molecular Medicine and Gene Therapy,Department of Laboratory Medicine,Faculty of Medicine
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Kohn, DB (author)
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Williams, D (author)
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von Kalle, C (author)
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(creator_code:org_t)
- 2003
- 2003
- English.
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In: Annals of the New York Academy of Sciences (HEMATOPOIETIC STEM CELLS 2002: GENETICS AND FUNCTION: Fourth International Symposium). - 0077-8923. ; 996, s. 112-121
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Abstract
Subject headings
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- The identification of unknown genomic flanking DNA sequences can be used for the molecular monitoring of retro-, lenti- and foamyviral integration, transgenes in early embryogenesis, insertional mutagenesis, cell fate, and stem cell plasticity. Most existing methods reflect shortcomings in sensitivity and or specificity, thus limiting genomic sequencing of unknown flanking DNA to clonal preparations. The application of linear amplification-mediated PCR (LAM-PCR), a recently developed direct sequencing technique for flanking DNA, should circumvent current limitations in different research fields. This technique combines preamplification of target DNA with a unique succession of enzymatic reactions on solid-phase. Using LAM-PCR, we show the previously unfeasible in vivo retro-, lenti- and foamyvirus integration site analysis in primate peripheral blood hematopoietic cells and human xenograft hematopoiesis. In light of two severe adverse events that occurred in a clinical SCID-X1 gene therapy trial, in vivo monitoring of the reinfused transduced cell pool by integration site analysis will be an important component of each gene transfer and therapy study aimed at clinical use.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Hematologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Hematology (hsv//eng)
Keyword
- proviruses
- therapy
- insertion sequence elements
- investigative techniques
- gene
- stem cells
Publication and Content Type
- kon (subject category)
- ref (subject category)
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- By the author/editor
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Schmidt, M
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Glimm, H
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Wissler, M
-
Hoffmann, G
-
Olsson, Karin
-
Sellers, S
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show more...
-
Carbonaro, D
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Tisdale, JF
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Leurs, C
-
Hanenberg, H
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Dunbar, CE
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Kiem, HP
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Karlsson, Stefan
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Kohn, DB
-
Williams, D
-
von Kalle, C
-
show less...
- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Clinical Medicin ...
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and Hematology
- Articles in the publication
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Annals of the Ne ...
- By the university
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Lund University