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A Split-Cre system designed to detect simultaneous expression of two genes based on SpyTag/SpyCatcher conjugation and Split-GFP dimerization

J Biol Chem. 2021-08; 
Xundong Wei, Jianhua Zhang, Jian Cui, Wei Xu, Xuyu Zhou, Jie Ma
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Custom Vector Construction SpyCatcher and SpyTag were directly synthesized based on the reported sequences (Integrated DNA Technologies and GenScript Biotech Corporation) (19) and subcloned into the pMSCV-IRES-Thy1.1 expression vector. Get A Quote

摘要

The Split-Cre system is a powerful tool for genetic manipulation and can be used to spatiotemporally control gene expression in vivo. However, the low activity of the reconstituted NCre/CCre recombinase in the Split-Cre system limits its application as an indicator of the simultaneous expression of a pair of genes of interest. Here, we describe two approaches for improving the activity of the Split-Cre system after Cre reconstitution based on self-associating split GFP (Split-GFP) and SpyTag/SpyCatcher conjugation. First, we created the Split-GFP-Cre system by constructing fusion proteins of NCre and CCre with the N-terminal and C-terminal subunits of GFP, respectively. Reconstitution of Cre by GFP-mediated di... More

关键词

Split-Cre, Split-GFP, Split-Spy-GCre, SpyTag/SpyCatcher