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This approach enabled hIL-7 to be expressed at consistent levels long-term, and improved human T lymphocyte numbers in the HIS model.
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The transgenes were expressed at consistent levels.
The ROSA26 locus was chosen for this study because it is widely thought to be expressed at a consistent level in mouse cells, and its use has been proven in numerous studies examining the effects of protein misexpression and as a Cre-inducible genetic tag (Zambrowicz et al., 1997; Kohlhepp et al., 2001; Safran et al., 2003).
Furthermore, these genes continue to be expressed at subsequent time points (E8.5, E9.5) consistent with a possible role in the maintenance or maturation of cardiac lineages.
Nevertheless, the proportion of OR genes considered to be expressed at detectable levels in this study is consistent with previous suggestions that up to 76% of OR genes are expressed at a detectable level in the human olfactory epithelium [ 10].
Consistent with this, the selenocysteine tRNA has also been shown to be expressed at all physiological/developmental stages (Figure S4 in [35]).
ORFan genes may be expressed at low levels (or infrequently), which may make them more tolerant of changes relative to genes expressed at high (and consistent) levels.
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Both fragments of sTEVp were cloned into a single vector (pBud-sol-sTEVp or pBud-mem-sTEVp; Fig. 5Ai), with each open reading frame driven by a different promoter, to ensure that the sTEVp fragments were expressed at a consistent ratio.
As expected, staining for BACE1 and BACE2 is absent from their respective knockouts while the complementary BACE is expressed at levels consistent with WT controls (Supporting Information Fig S3B).
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