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So the human genome evolved to shut down CMAH and Neu5Gc production.
To test the theory, the UCSD team compared rates of atherosclerosis in normal mice, which have a working CMAH gene, and mice genetically tweaked to have a switched-off gene.
Researchers have analyzed 322 animal genome sequences looking for animals with the presence of active CMAH genes.
At this point in human evolution, a certain gene, known as CMAH, that allows for the synthesis of a sugar called Neu5Gc, went missing.
(50) found that transcripts of CMAH, which generates Neu5Gc from Neu5Ac, are at very low levels in Western dog breed cells.
For detection of ZFN-induced mutations at the pig CMAH locus, the ZFN target locus was amplified from genomic DNA isolated from fibroblasts or cloned blastocysts (DNeasy kit, Qiagen) by nested PCR and subjected to the T7E1 assay (Kim et al., 2009).
To test the feasibility of using these knockout cell enrichment approaches for pig fibroblast cells, we used ZFNs targeting CMAH exon 6 (Figure 1a) and surrogate reporters containing either the eGFP gene alone (eGFP reporter) or eGFP and a truncated H-2Kk surface marker (MACS reporter), whose expression in cells can be recovered by a ZFN-induced frameshift mutation in CMAH.
After analyzing the effect of St presence on G, Ra,st, we assessed the effect of Cmah on the investigated responses.
Researchers compared two groups of mice: one with a functional CMAH gene, the other with an altered CMAH gene similar to the human mutation.
Clinical use of additional genetic modifications should be founded on further rigorous preclinical testing in NHPs to demonstrate their utility, which, in the case of CMAH, will likely require testing in an immunologically appropriate CMAH-KO large animal transplant model.
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