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Efficient characterization of retro-, lenti-, and foamyvector-transduced cell populations by high-accuracy insertion site sequencing

Schmidt, M; Glimm, H; Wissler, M; Hoffmann, G; Olsson, Karin LU ; Sellers, S; Carbonaro, D; Tisdale, JF; Leurs, C and Hanenberg, H, et al. (2003) HEMATOPOIETIC STEM CELLS 2002: GENETICS AND FUNCTION: Fourth International Symposium In Annals of the New York Academy of Sciences (HEMATOPOIETIC STEM CELLS 2002: GENETICS AND FUNCTION: Fourth International Symposium) 996. p.112-121
Abstract
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... (More)
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. (Less)
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Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
proviruses, therapy, insertion sequence elements, investigative techniques, gene, stem cells
in
Annals of the New York Academy of Sciences (HEMATOPOIETIC STEM CELLS 2002: GENETICS AND FUNCTION: Fourth International Symposium)
volume
996
pages
112 - 121
publisher
New York Academy of Sciences
conference name
HEMATOPOIETIC STEM CELLS 2002: GENETICS AND FUNCTION: Fourth International Symposium
external identifiers
  • wos:000183697800014
  • pmid:12799289
  • scopus:0037791981
ISSN
0077-8923
language
English
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yes
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ad87b2c1-e0e0-4be0-bbc6-117ff3d7d2f1 (old id 308144)
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http://www.annalsnyas.org/cgi/content/abstract/996/1/112
date added to LUP
2007-09-21 11:03:05
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2018-05-29 10:57:03
@inproceedings{ad87b2c1-e0e0-4be0-bbc6-117ff3d7d2f1,
  abstract     = {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.},
  author       = {Schmidt, M and Glimm, H and Wissler, M and Hoffmann, G and Olsson, Karin and Sellers, S and Carbonaro, D and Tisdale, JF and Leurs, C and Hanenberg, H and Dunbar, CE and Kiem, HP and Karlsson, Stefan and Kohn, DB and Williams, D and von Kalle, C},
  booktitle    = {Annals of the New York Academy of Sciences (HEMATOPOIETIC STEM CELLS 2002: GENETICS AND FUNCTION: Fourth International Symposium)},
  issn         = {0077-8923},
  keyword      = {proviruses,therapy,insertion sequence elements,investigative techniques,gene,stem cells},
  language     = {eng},
  pages        = {112--121},
  publisher    = {New York Academy of Sciences},
  title        = {Efficient characterization of retro-, lenti-, and foamyvector-transduced cell populations by high-accuracy insertion site sequencing},
  volume       = {996},
  year         = {2003},
}