Lentiviral and Retroviral Vector Construction

Lentiviral shRNA plasmids CCND1-1 (TRCN0000295875) and CCND1-2 (TRCN0000312873) were purchased from Sigma-Aldrich. The lentiviral shRNA vector for shRB1 was obtained from the RPCCC pGIPZ library (Clone ID: V2LHS_139). shRNAs targeting USP27X and ATXN7L3 were previously described (23 (link),28 (link)). Inducible shRNA vectors were generated by subcloning the USP27X targeting oligos (from the constitutive vectors described above) into the pLKO-Tet-On vector as described in (29 (link)). Retroviral vectors were generated by subcloning USP27X and CCND1 into a pBabe-Puro vector (Addgene) and lentiviral USP27X expressing vectors were generated by subcloning USP27X into a pLenti-Puro vector (Addgene).

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Alam S., Zunic A., Venkat S., Feigin M.E, & Atanassov B.S. (2022). Regulation of Cyclin D1 Degradation by Ubiquitin Specific Protease 27X is Critical for Cancer Cell Proliferation and Tumor Growth. Molecular cancer research : MCR, 20(12), 1751-1762.

Publication 2022

pBabe-Puro vector pLenti-Puro vector

Ccnd1 Clone Library Oligos Plasmids Retroviral Shrna Vector

Corresponding Organization : Roswell Park Comprehensive Cancer Center

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Variable analysis

independent variables

Lentiviral shRNA plasmids CCND1-1 (TRCN0000295875) and CCND1-2 (TRCN0000312873)
Lentiviral shRNA vector for shRB1 from the RPCCC pGIPZ library (Clone ID: V2LHS_139)
ShRNAs targeting USP27X and ATXN7L3
Inducible shRNA vectors generated by subcloning the USP27X targeting oligos into the pLKO-Tet-On vector
Retroviral vectors generated by subcloning USP27X and CCND1 into a pBabe-Puro vector
Lentiviral USP27X expressing vectors generated by subcloning USP27X into a pLenti-Puro vector

dependent variables

Unspecified

control variables

Unspecified

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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