Efficient characterization of retro-, lenti-, and foamyvector-transduced cell populations by high-accuracy insertion site sequencing
(2003) 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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/308144
- author
- organization
- publishing date
- 2003
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- subject
- keywords
- proviruses, therapy, insertion sequence elements, investigative techniques, gene, stem cells
- host publication
- 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
- conference dates
- 0001-01-02
- external identifiers
-
- wos:000183697800014
- pmid:12799289
- scopus:0037791981
- ISSN
- 0077-8923
- language
- English
- LU publication?
- yes
- id
- ad87b2c1-e0e0-4be0-bbc6-117ff3d7d2f1 (old id 308144)
- alternative location
- http://www.annalsnyas.org/cgi/content/abstract/996/1/112
- date added to LUP
- 2016-04-01 16:22:44
- date last changed
- 2022-01-28 19:17:45
@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}}, keywords = {{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}}, url = {{http://www.annalsnyas.org/cgi/content/abstract/996/1/112}}, volume = {{996}}, year = {{2003}}, }