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Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population

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Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population. / Kustikova, Olga S; Wahlers, Anke; Kuhlcke, Klaus; Stahle, Birgit; Zander, Axel R; Baum, Christopher; Fehse, Boris.

In: BLOOD, Vol. 102, No. 12, 01.12.2003, p. 3934-7.

Research output: SCORING: Contribution to journal › Short publication › Research › peer-review

Harvard

Kustikova, OS, Wahlers, A, Kuhlcke, K, Stahle, B, Zander, AR, Baum, C & Fehse, B 2003, 'Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population', BLOOD, vol. 102, no. 12, pp. 3934-7. https://doi.org/10.1182/blood-2003-05-1424

APA

Kustikova, O. S., Wahlers, A., Kuhlcke, K., Stahle, B., Zander, A. R., Baum, C., & Fehse, B. (2003). Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population. BLOOD, 102(12), 3934-7. https://doi.org/10.1182/blood-2003-05-1424

Vancouver

Kustikova OS, Wahlers A, Kuhlcke K, Stahle B, Zander AR, Baum C et al. Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population. BLOOD. 2003 Dec 1;102(12):3934-7. https://doi.org/10.1182/blood-2003-05-1424

Bibtex

@article{29bc9adb34b948afb384a68de4969426,
title = "Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population",
abstract = "Retroviral vectors are commonly used in clinical gene therapy, but recent observations of insertional oncogene activation in preclinical and clinical settings have forced a discussion of their safety. Here we investigated the relationship between retroviral transduction efficiency in mass cultures and the actual number of integrated vector copies in single cells using K562 leukemia and primary CD34+ cells. We found an exponential increase of integration numbers correlated to gene transfer rates and a linear increase of expression levels with insertion frequency. On average we detected one vector insertion per transduced cell for a gene transfer of less than 30%, 3 for 60%, and approximately 9 for 90% (in K562). Clonal analysis revealed strikingly increased variations of both transgene copy numbers (more than 20-fold in primary cells) and expression levels associated with higher transduction. Therefore, limiting retroviral gene transfer to approximately 30% may be suggested to avoid generating clones containing multiple insertions.",
keywords = "Cell Culture Techniques, Clone Cells, Flow Cytometry, Gene Dosage, Gene Expression, Genetic Vectors/administration & dosage, Green Fluorescent Proteins, Hematopoietic Stem Cells/metabolism, Humans, K562 Cells, Luminescent Proteins/analysis, Retroviridae/genetics, Transduction, Genetic/standards",
author = "Kustikova, {Olga S} and Anke Wahlers and Klaus Kuhlcke and Birgit Stahle and Zander, {Axel R} and Christopher Baum and Boris Fehse",
year = "2003",
month = dec,
day = "1",
doi = "10.1182/blood-2003-05-1424",
language = "English",
volume = "102",
pages = "3934--7",
journal = "BLOOD",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "12",

}

RIS

TY - JOUR

T1 - Dose finding with retroviral vectors: correlation of retroviral vector copy numbers in single cells with gene transfer efficiency in a cell population

AU - Kustikova, Olga S

AU - Wahlers, Anke

AU - Kuhlcke, Klaus

AU - Stahle, Birgit

AU - Zander, Axel R

AU - Baum, Christopher

AU - Fehse, Boris

PY - 2003/12/1

Y1 - 2003/12/1

N2 - Retroviral vectors are commonly used in clinical gene therapy, but recent observations of insertional oncogene activation in preclinical and clinical settings have forced a discussion of their safety. Here we investigated the relationship between retroviral transduction efficiency in mass cultures and the actual number of integrated vector copies in single cells using K562 leukemia and primary CD34+ cells. We found an exponential increase of integration numbers correlated to gene transfer rates and a linear increase of expression levels with insertion frequency. On average we detected one vector insertion per transduced cell for a gene transfer of less than 30%, 3 for 60%, and approximately 9 for 90% (in K562). Clonal analysis revealed strikingly increased variations of both transgene copy numbers (more than 20-fold in primary cells) and expression levels associated with higher transduction. Therefore, limiting retroviral gene transfer to approximately 30% may be suggested to avoid generating clones containing multiple insertions.

AB - Retroviral vectors are commonly used in clinical gene therapy, but recent observations of insertional oncogene activation in preclinical and clinical settings have forced a discussion of their safety. Here we investigated the relationship between retroviral transduction efficiency in mass cultures and the actual number of integrated vector copies in single cells using K562 leukemia and primary CD34+ cells. We found an exponential increase of integration numbers correlated to gene transfer rates and a linear increase of expression levels with insertion frequency. On average we detected one vector insertion per transduced cell for a gene transfer of less than 30%, 3 for 60%, and approximately 9 for 90% (in K562). Clonal analysis revealed strikingly increased variations of both transgene copy numbers (more than 20-fold in primary cells) and expression levels associated with higher transduction. Therefore, limiting retroviral gene transfer to approximately 30% may be suggested to avoid generating clones containing multiple insertions.

KW - Cell Culture Techniques

KW - Clone Cells

KW - Flow Cytometry

KW - Gene Dosage

KW - Gene Expression

KW - Genetic Vectors/administration & dosage

KW - Green Fluorescent Proteins

KW - Hematopoietic Stem Cells/metabolism

KW - Humans

KW - K562 Cells

KW - Luminescent Proteins/analysis

KW - Retroviridae/genetics

KW - Transduction, Genetic/standards

U2 - 10.1182/blood-2003-05-1424

DO - 10.1182/blood-2003-05-1424

M3 - Short publication

C2 - 12881303

VL - 102

SP - 3934

EP - 3937

JO - BLOOD

JF - BLOOD

SN - 0006-4971

IS - 12

ER -