Methods of Titrating Nonselectable Recombinant Retroviruses
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Last Update: 2021-02-24
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Source: Internet
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Author: User
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When using recombinant retroviruses for gene transduction, it is necessary to have a measurement of the concentration of virus particles in the medium conditioned by virus-producing cells (i.e., the viral titer). In the case of viruses encoding selectable markers, this can readily be determined by infecting a susceptible cell line (usually 3T3 fibroblasts), culturing in selection medium, and scoring colonies of resistant cells (Chapter 3 , this volume), which gives a concentration of virus particles in the preparation as a number of colony-forming units/mL (CFU/mL). In cases in which the gene of interest is not selectable, quantitation can still be achieved by the use of recombinant viruses additionally encoding a selectable marker under the control of a separate promoter (Chapter 1 ). In some cases a selectable gene may be desirable, but there is evidence indicating that in viruses possessing two transcriptional units expression from each is depressed (
1
,
2
). Thus, for maximum viral titer and maximum gene expression in the target cell, the virus construct of choice will encode a single gene. Other means of estimating viral titers are then required.
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