Abstract
RCAS (B/P) is a replication-competent avian retrovirus engineered by Hughes et al. (J Virol 61:3004–3012, 1987) and is referred to in this chapter as RCAS for simplicity. The RCAS retrovirus has been widely used as a vehicle for stable transduction of a gene into cells both in the developing chick embryo and tissue/cell culture. It can be used for both gain- and loss-function experiments. The ability of this virus to spread among proliferating cells makes it possible to achieve widespread gene transduction in the developing retina. The transduction efficiency of RCAS is highly depending on the titer of the viral stock, particularly for experiments involving solid tissues such as the developing retina. Here, we describe the procedure that we have used for 15 years to generate RCAS viral stocks with a titer of 1–5 × 108 pfu/ml.
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References
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Acknowledgments
This work is supported by NIH/NEI EY011640, EyeSight Foundation of Alabama FY2011-12-276, and an unrestricted grant to UAB Department of Ophthalmology from Research to Prevent Blindness.
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Yan, RT., Wang, SZ. (2012). Production of High-Titer RCAS Retrovirus. In: Wang, SZ. (eds) Retinal Development. Methods in Molecular Biology, vol 884. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-848-1_13
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DOI: https://doi.org/10.1007/978-1-61779-848-1_13
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-61779-848-1
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