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Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR

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Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR. / Christodoulou, I.; Patsali, P.; Stephanou, C.; Antoniou, M.; Kleanthous, M.; Lederer, C. W.

In: Gene Therapy, Vol. 23, No. 1, 01.01.2016, p. 113-118.

Research output: Contribution to journalArticle

Harvard

Christodoulou, I, Patsali, P, Stephanou, C, Antoniou, M, Kleanthous, M & Lederer, CW 2016, 'Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR', Gene Therapy, vol. 23, no. 1, pp. 113-118. https://doi.org/10.1038/gt.2015.60

APA

Christodoulou, I., Patsali, P., Stephanou, C., Antoniou, M., Kleanthous, M., & Lederer, C. W. (2016). Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR. Gene Therapy, 23(1), 113-118. https://doi.org/10.1038/gt.2015.60

Vancouver

Christodoulou I, Patsali P, Stephanou C, Antoniou M, Kleanthous M, Lederer CW. Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR. Gene Therapy. 2016 Jan 1;23(1):113-118. https://doi.org/10.1038/gt.2015.60

Author

Christodoulou, I. ; Patsali, P. ; Stephanou, C. ; Antoniou, M. ; Kleanthous, M. ; Lederer, C. W. / Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR. In: Gene Therapy. 2016 ; Vol. 23, No. 1. pp. 113-118.

Bibtex Download

@article{18526c1629d94422b4aa5a56ec3c642f,
title = "Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR",
abstract = "Lentiviruses are the vectors of choice for many preclinical studies and clinical applications of gene therapy. Accurate measurement of biological vector titre before treatment is a prerequisite for vector dosing, and the calculation of vector integration sites per cell after treatment is as critical to the characterisation of modified cell products as it is to long-term follow-up and the assessment of risk and therapeutic efficiency in patients. These analyses are typically based on quantitative real-time PCR (qPCR), but as yet compromise accuracy and comparability between laboratories and experimental systems, the former by using separate simplex reactions for the detection of endogene and lentiviral sequences and the latter by designing different PCR assays for analyses in human cells and animal disease models. In this study, we validate in human and murine cells a qPCR system for the single-tube assessment of lentiviral vector copy numbers that is suitable for analyses in at least 33 different mammalian species, including human and other primates, mouse, pig, cat and domestic ruminants. The established assay combines the accuracy of single-tube quantitation by duplex qPCR with the convenience of one-off assay optimisation for cross-species analyses and with the direct comparability of lentiviral transduction efficiencies in different species.",
author = "I. Christodoulou and P. Patsali and C. Stephanou and M. Antoniou and M. Kleanthous and Lederer, {C. W.}",
year = "2016",
month = jan,
day = "1",
doi = "10.1038/gt.2015.60",
language = "English",
volume = "23",
pages = "113--118",
journal = "Gene Therapy",
issn = "0969-7128",
publisher = "Nature Publishing Group",
number = "1",

}

RIS (suitable for import to EndNote) Download

TY - JOUR

T1 - Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR

AU - Christodoulou, I.

AU - Patsali, P.

AU - Stephanou, C.

AU - Antoniou, M.

AU - Kleanthous, M.

AU - Lederer, C. W.

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Lentiviruses are the vectors of choice for many preclinical studies and clinical applications of gene therapy. Accurate measurement of biological vector titre before treatment is a prerequisite for vector dosing, and the calculation of vector integration sites per cell after treatment is as critical to the characterisation of modified cell products as it is to long-term follow-up and the assessment of risk and therapeutic efficiency in patients. These analyses are typically based on quantitative real-time PCR (qPCR), but as yet compromise accuracy and comparability between laboratories and experimental systems, the former by using separate simplex reactions for the detection of endogene and lentiviral sequences and the latter by designing different PCR assays for analyses in human cells and animal disease models. In this study, we validate in human and murine cells a qPCR system for the single-tube assessment of lentiviral vector copy numbers that is suitable for analyses in at least 33 different mammalian species, including human and other primates, mouse, pig, cat and domestic ruminants. The established assay combines the accuracy of single-tube quantitation by duplex qPCR with the convenience of one-off assay optimisation for cross-species analyses and with the direct comparability of lentiviral transduction efficiencies in different species.

AB - Lentiviruses are the vectors of choice for many preclinical studies and clinical applications of gene therapy. Accurate measurement of biological vector titre before treatment is a prerequisite for vector dosing, and the calculation of vector integration sites per cell after treatment is as critical to the characterisation of modified cell products as it is to long-term follow-up and the assessment of risk and therapeutic efficiency in patients. These analyses are typically based on quantitative real-time PCR (qPCR), but as yet compromise accuracy and comparability between laboratories and experimental systems, the former by using separate simplex reactions for the detection of endogene and lentiviral sequences and the latter by designing different PCR assays for analyses in human cells and animal disease models. In this study, we validate in human and murine cells a qPCR system for the single-tube assessment of lentiviral vector copy numbers that is suitable for analyses in at least 33 different mammalian species, including human and other primates, mouse, pig, cat and domestic ruminants. The established assay combines the accuracy of single-tube quantitation by duplex qPCR with the convenience of one-off assay optimisation for cross-species analyses and with the direct comparability of lentiviral transduction efficiencies in different species.

UR - http://www.scopus.com/inward/record.url?scp=84955182083&partnerID=8YFLogxK

U2 - 10.1038/gt.2015.60

DO - 10.1038/gt.2015.60

M3 - Article

C2 - 26202078

AN - SCOPUS:84955182083

VL - 23

SP - 113

EP - 118

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 1

ER -

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