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231226s2023 xx |||||o 00| ||eng c |
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|a 10.1002/adma.202209904
|2 doi
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|a pubmed24n1395.xml
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|a (DE-627)NLM353150029
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|a (NLM)36808641
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|a DE-627
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|e rakwb
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|a eng
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|a Bretherton, Ross C
|e verfasserin
|4 aut
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|a User-Controlled 4D Biomaterial Degradation with Substrate-Selective Sortase Transpeptidases for Single-Cell Biology
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|c 2023
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
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|2 rdamedia
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|a ƒa Online-Ressource
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|2 rdacarrier
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|a Date Completed 12.05.2023
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|a Date Revised 02.05.2024
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a © 2023 Wiley-VCH GmbH.
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|a Stimuli-responsive biomaterials show great promise for modeling disease dynamics ex vivo with spatiotemporal control over the cellular microenvironment. However, harvesting cells from such materials for downstream analysis without perturbing their state remains an outstanding challenge in 3/4-dimensional (3D/4D) culture and tissue engineering. In this manuscript, a fully enzymatic strategy for hydrogel degradation that affords spatiotemporal control over cell release while maintaining cytocompatibility is introduced. Exploiting engineered variants of the sortase transpeptidase evolved to recognize and selectively cleave distinct peptide sequences largely absent from the mammalian proteome, many limitations implicit to state-of-the-art methods to liberate cells from gels are sidestepped. It is demonstrated that evolved sortase exposure has minimal impact on the global transcriptome of primary mammalian cells and that proteolytic cleavage proceeds with high specificity; incorporation of substrate sequences within hydrogel crosslinkers permits rapid and selective cell recovery with high viability. In composite multimaterial hydrogels, it is shown that sequential degradation of hydrogel layers enables highly specific retrieval of single-cell suspensions for phenotypic analysis. It is expected that the high bioorthogonality and substrate selectivity of the evolved sortases will lead to their broad adoption as an enzymatic material dissociation cue and that their multiplexed use will enable newfound studies in 4D cell culture
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|a Journal Article
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|a biomaterials
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|a bioorthogonal chemistry
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|a degradation
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|a hydrogels
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|a single-cell biology
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|a sortase
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|a Biocompatible Materials
|2 NLM
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|a Peptidyl Transferases
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|a EC 2.3.2.12
|2 NLM
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|a Peptides
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|a Hydrogels
|2 NLM
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|a Haack, Amanda J
|e verfasserin
|4 aut
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1 |
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|a Kopyeva, Irina
|e verfasserin
|4 aut
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|a Rahman, Fariha
|e verfasserin
|4 aut
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|a Kern, Jonah D
|e verfasserin
|4 aut
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|a Bugg, Darrian
|e verfasserin
|4 aut
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1 |
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|a Theberge, Ashleigh B
|e verfasserin
|4 aut
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|a Davis, Jennifer
|e verfasserin
|4 aut
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| 700 |
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|a DeForest, Cole A
|e verfasserin
|4 aut
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| 773 |
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|i Enthalten in
|t Advanced materials (Deerfield Beach, Fla.)
|d 1998
|g 35(2023), 19 vom: 01. Mai, Seite e2209904
|w (DE-627)NLM098206397
|x 1521-4095
|7 nnns
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| 773 |
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|g volume:35
|g year:2023
|g number:19
|g day:01
|g month:05
|g pages:e2209904
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|u http://dx.doi.org/10.1002/adma.202209904
|3 Volltext
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