Engineered Telomere Enrichment Vectors

The ReDACT-TR technique was motivated by the use of a telomere seed sequence to generate monosomic cells as described in ref^{1 (link)}. To enhance the efficacy of this approach, we created new plasmids linking a telomere seed sequence with a puromycin selection marker, which allowed us to enrich for stably-transfected aneuploidy-loss cells using drug selection. These EF1a-Puro-Telo vectors (Addgene #195138 and #195139) were generated by introducing a puromycin selection marker to a telomere seed sequence gifted by Alison Taylor. This vector was digested with FastDigest KpnI (Thermo Fisher Scientific, cat. no. FD0524) and FastDigest BstZ17I (Thermo Fisher Scientific, cat. no. FD0704), and gel purified to obtain the artificial telomere construct.

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Girish V., Lakhani A.A., Scaduto C.M., Thompson S.L., Brown L.M., Hagenson R.A., Sausville E.L., Mendelson B.E., Lukow D.A., Yuan M.L., Kandikuppa P.K., Stevens E.C., Lee S.N., Salovska B., Li W., Smith J.C., Taylor A.M., Martienssen R.A., Liu Y., Sun R, & Sheltzer J.M. (2023). Oncogene-like addiction to aneuploidy in human cancers. bioRxiv.

Publication Preprint 2023

FastDigest KpnI

Cells Cells aneuploidy Drug Plasmids Puromycin Ref1 Telomere Vector

Corresponding Organization : Yale University

Other organizations : Johns Hopkins Medicine, Johns Hopkins University, Cold Spring Harbor Laboratory, Columbia University, Howard Hughes Medical Institute, University of Minnesota

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Variable analysis

independent variables

Introducing a puromycin selection marker to a telomere seed sequence

dependent variables

Enrichment for stably-transfected aneuploidy-loss cells using drug selection

control variables

Telomere seed sequence gifted by Alison Taylor

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