Part:BBa_K4469002

WPRE is a DNA sequence motif that enhances gene expression by forming a tertiary structure post-transcription. It is a cis-acting component from Woodchuck Hepatitis Virus (WHV) genome (1093-1684bp) (Ong et al., 2017). Containing gamma, alpha and beta elements, insertion of WPRE in 3’ untranslated region (3’ UTR) of the coding sequence has improved gene delivery by oncoretroviral and lentiviral vectors. Green fluorescent protein (GFP) and luciferase signals have shown to be 5- to 8-fold higher than control with WPRE placed downstream (Donello et al., 1998).

As WPRE enhances the expression of a transgene lacking introns, it is frequently used in molecular cloning to enhance transcription of genes delivered by viral vectors (Donello et al., 1998). It has shown to increase mRNA activity by several folds in several mouse and human primary cell lines, and in vivo in mouse liver (Zufferey et al., 1999). The 589nt-long sequence also contributes to strong expression of transgene in AAV system. WPRE resulted in efficient GFP expression in striatal neurons (Paterna et al., 2000); enhanced RFP expression in human kidney, mouse myoblast, and human neuroblastoma cells (Wang et al., 2016); 2-50 fold heightened luciferase expression in mouse liver, kidney and lung (Xu et al., 2004), all demonstrated its ability to increase transgene expression in a non-cell-type and non-transgene specific manner when included in an AAV vector.

Note: WPRE can only exert its influence when placed in the sense orientation (Donello et al., 1998).

Reference

Donello, J. E., Loeb, J. E., & Hope, T. J. (1998). Woodchuck hepatitis virus contains a tripartite posttranscriptional regulatory element. Journal of Virology, 72(6), 5085–5092. https://doi.org/10.1128/jvi.72.6.5085-5092.1998

Ong, J. M., Brown, C. R., Mendel, M. C., & Cost, G. J. (2017). The WPRE improves genetic engineering with site-specific nucleases. https://doi.org/10.1101/126904

Paterna, J.-C., Moccetti, T., Mura, A., Feldon, J., & Büeler, H. (2000). Influence of promoter and WHV post-transcriptional regulatory element on AAV-mediated transgene expression in The rat brain. Gene Therapy, 7(15), 1304–1311. https://doi.org/10.1038/sj.gt.3301221

Wang, L., Wang, Z., Zhang, F., Zhu, R., Bi, J., Wu, J., Zhang, H., Wu, H., Kong, W., Yu, B., & Yu, X. (2016). Enhancing transgene expression from recombinant AAV8 vectors in different tissues using woodchuck hepatitis virus post-transcriptional regulatory element. International Journal of Medical Sciences, 13(4), 286–291. https://doi.org/10.7150/ijms.14152

Xu, Z., Yue, Y., Lai, Y., Ye, C., Qiu, J., Pintel, D. J., & Duan, D. (2004). Trans-splicing adeno-associated viral vector-mediated gene therapy is limited by the accumulation of spliced mrna but not by dual vector coinfection efficiency. Human Gene Therapy, 15(9), 896–905. https://doi.org/10.1089/hum.2004.15.896

Zufferey, R., Donello, J. E., Trono, D., & Hope, T. J. (1999). Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors. Journal of Virology, 73(4), 2886–2892. https://doi.org/10.1128/jvi.73.4.2886-2892.1999