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Phoenix a amphotropic packaging cells

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Phoenix A amphotropic packaging cells are a cell line designed for the production of retroviral vectors. These cells are capable of packaging retroviral genomes into infectious viral particles that can be used for the delivery of genetic material to target cells.

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Phoenix amphotropic cells Phoenix packaging cells Phoenix cells Phoenix A

2 protocols using phoenix a amphotropic packaging cells

1

Overexpression of NLSTBLR1 in LNCaP Cells

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We created a TBLR1 fusion protein in which the strong NLS PKKKRKV was fused to the N terminus of TBLR1 to create NLSTBLR1. A retroviral-based mammalian expression vector, pBabe, was used to stably express NLSTBLR1 proteins in LNCaP cells as described previously (Peng et al. 2008a (link)). Briefly, Phoenix A amphotropic packaging cells (American Type Culture Collection) were transfected with NLSTBLR1, and pBabe retroviral constructs to produce virus. The virus-containing supernatant was collected by centrifugation and filtered before retroviral infection. Stable cell lines were selected in 1μg/mL puromycin.
Daniels G., Li Y., Gellert L.L., Zhou A., Melamed J., Wu X., Zhang X., Zhang D., Meruelo D., Logan S.K., Basch R, & Lee P. (2014). TBLR1 as an AR coactivator selectively activates AR target genes to inhibit prostate cancer growth. Endocrine-related cancer, 21(1), 127-142.
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2

Engineered TBLR1 Fusion Proteins

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A TBLR1 fusion protein was created in which the strong NES (MLQKKLEELE) was fused to the N terminus of TBLR1 to create NESTBLR1. An additional fusion protein was created in which the strong NES was fused to the sequence of AA89-514 from full length TBLR1 to create NEScvTBLR1. Both fusion proteins were cloned into the pBabe Puro retroviral vector as described previously [19 (link)]. Briefly, Phoenix A amphotropic packaging cells (American Type Culture Collection) were transfected with NESTBLR1, NEScvTBLR1 and pBabe retroviral constructs to produce virus. The virus-containing supernatant was collected by centrifugation and filtered before retroviral infection. Stable cell lines were selected in 1 μg/mL puromycin.
Daniels G., Zhang X., Zhong X., Santiago L., Wang L.H., Wu X., Zhang J.Y., Liang F., Li X., Neubert T.A., Steinke L., Shen Y., Basch R., Schneider R., Levy D.E, & Lee P. (2016). Cytoplasmic, full length and novel cleaved variant, TBLR1 reduces apoptosis in prostate cancer under androgen deprivation. Oncotarget, 7(26), 39556-39571.
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