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Induction of endogenous Bcl-xS through the control of Bcl-x pre-mRNA splicing by antisense oligonucleotides

Abstract

Resistance to apoptosis, which plays an important role in tumors that are refractory to chemotherapy, is regulated by the ratio of antiapoptotic to proapoptotic proteins. By manipulating levels of these proteins, cells can become sensitized to undergo apoptosis in response to chemotherapeutic agents. Alternative splicing of the bcl-x gene gives rise to two proteins with antagonistic functions: Bcl-xL, a well-characterized antiapoptotic protein, and Bcl-xS, a proapoptotic protein. We show here that altering the ratio of Bcl-xL to Bcl-xS in the cell using an antisense oligonucleotide permitted cells to be sensitized to undergo apoptosis in response to ultraviolet B radiation and chemotherapeutic drug treatment. These results demonstrate the ability of a chemically modified oligonucleotide to alter splice site selection in an endogenous gene and illustrate a powerful tool to regulate cell survival.

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Figure 1: (A) Illustration of the bcl-x gene and alternative splice products.
Figure 2: RPA analysis of oligonucleotide effects on Bcl-x splicing.
Figure 3: Effect of oligonucleotide concentration on Bcl-x expression levels.
Figure 4: Analysis of Bcl-x protein levels.
Figure 5: Physiological effect of Bcl-xS induction.

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Acknowledgements

We would like to thank Sue Freier, Todd Burckin, and Frank Bennett for advice with these studies.

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Correspondence to Nicholas M. Dean.

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Taylor, J., Zhang, Q., Wyatt, J. et al. Induction of endogenous Bcl-xS through the control of Bcl-x pre-mRNA splicing by antisense oligonucleotides. Nat Biotechnol 17, 1097–1100 (1999). https://doi.org/10.1038/15079

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