Generating stable WRN-deficient cell lines

The SV40-transformed WRN-deficient fibroblast cell line (AG11395) was obtained from Coriell Cell Repositories (Camden, NJ, USA). To produce stable cell lines, AG11395 (WS) fibroblasts were transduced with retroviruses expressing the full-length cDNA encoding wild-type WRN (WS^WT), exonuclease-dead (WS^E84A) or helicase-dead (WS^K577M) (21 (link)). All the cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM; Life Technologies) supplemented with 10% FBS (Boehringer Mannheim) and incubated at 37°C in a humidified 5% CO₂ atmosphere.

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Aiello F.A., Palma A., Malacaria E., Zheng L., Campbell J.L., Shen B., Franchitto A, & Pichierri P. (2019). RAD51 and mitotic function of mus81 are essential for recovery from low-dose of camptothecin in the absence of the WRN exonuclease. Nucleic Acids Research, 47(13), 6796-6810.

Publication 2019

AG11395 Dulbecco's modified Eagle's medium FBS

Atmosphere Cdna Cell Cell lines Exonuclease Fibroblasts Helicase Retroviruses Sv40 Transformed cell line

Corresponding Organization : Istituto Nazionale Biostrutture e Biosistemi

Other organizations : City of Hope, California Institute of Technology

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Variable analysis

independent variables

Expression of the full-length cDNA encoding wild-type WRN (WS^WT^)
Expression of exonuclease-dead (WS^E84A^)
Expression of helicase-dead (WS^K577M^)

dependent variables

Not explicitly mentioned

control variables

AG11395 (WS) fibroblasts

controls

AG11395 (WS) fibroblasts as a negative control

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