ACS polymers AuPub Date : 2024-12-20eCollection Date: 2025-02-12DOI: 10.1021/acspolymersau.4c00085
Hugo J Rodríguez-Franco, Pauline B M Hendrickx, Maartje M C Bastings
{"title":"Tailoring DNA Origami Protection: A Study of Oligolysine-PEG Coatings for Improved Colloidal, Structural, and Functional Integrity.","authors":"Hugo J Rodríguez-Franco, Pauline B M Hendrickx, Maartje M C Bastings","doi":"10.1021/acspolymersau.4c00085","DOIUrl":"10.1021/acspolymersau.4c00085","url":null,"abstract":"<p><p>Application of protective polymer coatings to enhance the biostability of DNA-based nanomaterials (DONs) has become common practice in <i>in vitro</i> and <i>in vivo</i> experiments. While the functional effect of these coatings is obvious, a detailed molecular picture of what is protected and for how long remains unclear. Additionally, the use of the oligolysine-1kPEG protective polymer has been limited due to aggregation issues. In this study, we evaluated the colloidal stability, structural integrity, and functional preservation of DONs coated with oligolysine (K)-1k/5kPEG block copolymers. Dynamic light scattering and transmission electron microscopy were employed to assess colloidal stability before and after degradation. A FRET-based assay was developed to monitor the directionality of degradation, while quantitative PCR measured the protection of functional DNA handles, crucial for the design of ligand-functionalized DONs. Our results show that K<sub>10</sub>-1kPEG, while prone to aggregation, can offer similar protection against nucleases as K<sub>10</sub>-5kPEG, provided buffer conditions are carefully chosen. Maintaining the colloidal, structural, and functional stability before and after nuclease exposure supports DON applications, particularly at the biointerface. These insights provide valuable guidelines for selecting the most effective protection strategy and enhancing DON functionality across diverse biological environments.</p>","PeriodicalId":72049,"journal":{"name":"ACS polymers Au","volume":"5 1","pages":"35-44"},"PeriodicalIF":4.7,"publicationDate":"2024-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11826485/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143434343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS polymers AuPub Date : 2024-12-19DOI: 10.1021/acspolymersau.4c0008510.1021/acspolymersau.4c00085
Hugo J. Rodríguez-Franco, Pauline B. M. Hendrickx and Maartje M. C. Bastings*,
{"title":"Tailoring DNA Origami Protection: A Study of Oligolysine-PEG Coatings for Improved Colloidal, Structural, and Functional Integrity","authors":"Hugo J. Rodríguez-Franco, Pauline B. M. Hendrickx and Maartje M. C. Bastings*, ","doi":"10.1021/acspolymersau.4c0008510.1021/acspolymersau.4c00085","DOIUrl":"https://doi.org/10.1021/acspolymersau.4c00085https://doi.org/10.1021/acspolymersau.4c00085","url":null,"abstract":"<p >Application of protective polymer coatings to enhance the biostability of DNA-based nanomaterials (DONs) has become common practice in <i>in vitro</i> and <i>in vivo</i> experiments. While the functional effect of these coatings is obvious, a detailed molecular picture of what is protected and for how long remains unclear. Additionally, the use of the oligolysine-1kPEG protective polymer has been limited due to aggregation issues. In this study, we evaluated the colloidal stability, structural integrity, and functional preservation of DONs coated with oligolysine (K)-1k/5kPEG block copolymers. Dynamic light scattering and transmission electron microscopy were employed to assess colloidal stability before and after degradation. A FRET-based assay was developed to monitor the directionality of degradation, while quantitative PCR measured the protection of functional DNA handles, crucial for the design of ligand-functionalized DONs. Our results show that K<sub>10</sub>-1kPEG, while prone to aggregation, can offer similar protection against nucleases as K<sub>10</sub>-5kPEG, provided buffer conditions are carefully chosen. Maintaining the colloidal, structural, and functional stability before and after nuclease exposure supports DON applications, particularly at the biointerface. These insights provide valuable guidelines for selecting the most effective protection strategy and enhancing DON functionality across diverse biological environments.</p>","PeriodicalId":72049,"journal":{"name":"ACS polymers Au","volume":"5 1","pages":"35–44 35–44"},"PeriodicalIF":4.7,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acspolymersau.4c00085","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143385978","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS polymers AuPub Date : 2024-12-16DOI: 10.1021/acspolymersau.4c0007310.1021/acspolymersau.4c00073
Kayo Terada, Kousuke Tsuchiya, Alexandros Lamprou and Keiji Numata*,
{"title":"Chemoenzymatic Synthesis of Poly-l-lysine via Esterification with Alcohol in One-Pot","authors":"Kayo Terada, Kousuke Tsuchiya, Alexandros Lamprou and Keiji Numata*, ","doi":"10.1021/acspolymersau.4c0007310.1021/acspolymersau.4c00073","DOIUrl":"https://doi.org/10.1021/acspolymersau.4c00073https://doi.org/10.1021/acspolymersau.4c00073","url":null,"abstract":"<p >Chemoenzymatic polymerization (CEP) using enzymes as catalysts is gaining attention as an environmentally friendly method for synthesizing polypeptides. This method proceeds under mild conditions in aqueous solvents and leverages the substrate specificity of enzymes, allowing polymerization reactions to occur without the need to protect reactive side-chain functional groups. However, the monomers used must have esterified C-termini, such as amino acids or oligopeptides. In this study, we used <span>l</span>-lysine (Lys-OH) as a model example and performed one-pot CEP with papain without isolating the esterified lysine. Esterification of Lys-OH was achieved by using hydrochloric acid as a catalyst in ethanol, and one-pot polymerization resulted in poly-<span>l</span>-lysine (polyLys) with a peak top degree of polymerization (DP) of 6 and a maximum DP of 18, with a 31% conversion from the nonesterified lysine. The obtained polyLys was all α-linked, demonstrating that regioselective polymerization was successfully achieved even with one-pot CEP.</p>","PeriodicalId":72049,"journal":{"name":"ACS polymers Au","volume":"5 1","pages":"26–34 26–34"},"PeriodicalIF":4.7,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acspolymersau.4c00073","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143385975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Chemoenzymatic Synthesis of Poly-l-lysine via Esterification with Alcohol in One-Pot.","authors":"Kayo Terada, Kousuke Tsuchiya, Alexandros Lamprou, Keiji Numata","doi":"10.1021/acspolymersau.4c00073","DOIUrl":"10.1021/acspolymersau.4c00073","url":null,"abstract":"<p><p>Chemoenzymatic polymerization (CEP) using enzymes as catalysts is gaining attention as an environmentally friendly method for synthesizing polypeptides. This method proceeds under mild conditions in aqueous solvents and leverages the substrate specificity of enzymes, allowing polymerization reactions to occur without the need to protect reactive side-chain functional groups. However, the monomers used must have esterified C-termini, such as amino acids or oligopeptides. In this study, we used l-lysine (Lys-OH) as a model example and performed one-pot CEP with papain without isolating the esterified lysine. Esterification of Lys-OH was achieved by using hydrochloric acid as a catalyst in ethanol, and one-pot polymerization resulted in poly-l-lysine (polyLys) with a peak top degree of polymerization (DP) of 6 and a maximum DP of 18, with a 31% conversion from the nonesterified lysine. The obtained polyLys was all α-linked, demonstrating that regioselective polymerization was successfully achieved even with one-pot CEP.</p>","PeriodicalId":72049,"journal":{"name":"ACS polymers Au","volume":"5 1","pages":"26-34"},"PeriodicalIF":4.7,"publicationDate":"2024-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11826487/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143434333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}