Doxycycline-Inducible Polycistronic Reprogramming Vector

The doxycycline-inducible polycistronic reprogramming vector was generated by cloning a GFP-IRES sequence downstream of the tetracycline response element in the backbone FUW-tetO-hOKMS (Addgene 51543, a gift from Cacchiarelli et al., 2015 (link)). This vector was used in combination with FUW-M2rtTA (Addgene 20342, a gift from Hockemeyer et al., 2008 (link)). Viral particles were generated by transfecting HEK293T cells with the packaging plasmids pMD2.G (Addgene 12259, a gift from Didier Trono) and psPAX2 (Addgene 12260, a gift from Didier Trono) and either FUW-tetO-GFP-hOKMS or FUW-M2rtTA.

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Gill D., Parry A., Santos F., Okkenhaug H., Todd C.D., Hernando-Herraez I., Stubbs T.M., Milagre I, & Reik W. (2022). Multi-omic rejuvenation of human cells by maturation phase transient reprogramming. eLife, 11, e71624.

Publication 2022

FUW-tetO-hOKMS FUW-M2rtTA pMD2.G psPAX2 FUW-tetO-GFP-hOKMS

Backbone Cells Doxycycline Ires Plasmids Response element Tetracycline Vector Viral particles

Corresponding Organization :

Other organizations : Babraham Institute, Instituto Gulbenkian de Ciência, Wellcome Sanger Institute, University of Cambridge

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Variable analysis

independent variables

Doxycycline-inducible polycistronic reprogramming vector
FUW-M2rtTA vector

dependent variables

Viral particle generation

control variables

Packaging plasmids pMD2.G and psPAX2

controls

Positive control: FUW-tetO-GFP-hOKMS vector
Negative control: FUW-M2rtTA vector

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