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These data demonstrate that ZFN-mediated gene editing is a highly effective tool for dissection of pre-mRNA splicing regulatory sequences in their endogenous context.
For example TSSs 4 and 5 of PSMD8 are stronger in the heterologous than the endogenous context, and the major TSS 3 of endogenous WBP11 is weaker in the heterologous context.
The red color scale depicts the DHS signal within the fragments in their endogenous context.
We next sought to address if stable-bleb cells also emerge in the endogenous context of embryonic development.
22 Albeit elaborate, the proposed approach on predicting host parasite interactions lacks stringent evaluation of the predicted protein partners in endogenous context.
These results demonstrate that the 874 bp apRXa wing enhancer element is required but not sufficient in the endogenous context to correctly regulate ap expression.
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During human endotoxemia and sepsis, circulating Angpt1 levels remain unchanged or even decrease, whereas the endogenous context-specific Tie-2 antagonist, angiopoietin-2 (Angpt2), is rapidly released by the activated endothelium and disrupts the constitutive Angpt1/Tie2 signaling by preventing Angpt1 from binding to the receptor [ 12- 19].
We performed ZFN-mediated targeted mutagenesis of the C. griseus BAX gene to dissect the importance of various splice site sequences in their normal, endogenous chromosomal context.
To examine the expression of antagonistic ILPs in a more endogenous genomic context, we generated functional fosmid INS-18 and INS-1 reporters.
This has many applications in gene deletion, analysis of defined genetic changes, visualisation, purification and expression analysis of genes or their protein products in an endogenous cellular context that retains all of the regulatory elements present within the genome.
However, as we initially employed a wild-type S. typhimurium strain expressing enhanced levels of SipA from a plasmid (sipA ++; Cain et al., 2004), we next examined the localization of SipAFLAG expressed from the endogenous chromosomal context.
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