期刊
ACS SYNTHETIC BIOLOGY
卷 8, 期 10, 页码 2315-2326出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.9b00127
关键词
mucin; glycan; glycosylation; synthetic biology; engineering; custom gene synthesis
资金
- National Institute of General Medical Sciences Ruth L. Kirschstein National Research Service Award [2T32GM008267]
- National Science Foundation Graduate Research Fellowship [DGE-1650441]
- National Institute of Health [DP2 GM229133]
- National Cancer Institute [U54 CA210184]
- National Science Foundation [1752226]
- NYSTEM Grant [C029155]
- NIH [S10OD018516]
- National Institutes of Health [1S10OD018530, P41GM10349010]
- Directorate For Engineering
- Div Of Civil, Mechanical, & Manufact Inn [1752226] Funding Source: National Science Foundation
Few approaches exist for the stable and controllable synthesis of customized mucin glycoproteins for glycocalyx editing in eukaryotic cells. Taking advantage of custom gene synthesis and a biology-by-parts approach to cDNA construction, we build a library of swappable DNA bricks for mucin leader tags, membrane anchors, cytoplasmic motifs, and optical reporters, as well as codon-optimized native mucin repeats and newly designed domains for synthetic mucins. We construct a library of over 50 mucins, each with unique chemical, structural, and optical properties and describe how additional permutations could readily be constructed. We apply the library to explore sequence-specific effects on glycosylation for engineering of mucins. We find that the extension of the immature alpha-GalNAc Tn-antigen to Core 1 and Core 2 glycan structures depends on the underlying peptide backbone sequence. Glycosylation could also be influenced through recycling motifs on the mucin cytoplasmic tail. We expect that the mucin parts inventory presented here can be broadly applied for glycocalyx research and mucin-based biotechnologies.
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