Show simple item record

dc.contributor.authorGayet, Raphael V.
dc.contributor.authorde Puig Guixe, Helena
dc.contributor.authorEnglish, Max A.
dc.contributor.authorSoenksen Martinez, Luis Ruben
dc.contributor.authorNguyen, Peter Q.
dc.contributor.authorMao, Angelo S.
dc.contributor.authorAngenent-Mari, Nicolaas M.
dc.contributor.authorCollins, James J.
dc.date.accessioned2021-09-21T19:25:52Z
dc.date.available2021-09-21T19:25:52Z
dc.date.issued2020-08
dc.date.submitted2019-12
dc.identifier.issn1754-2189
dc.identifier.issn1750-2799
dc.identifier.urihttps://hdl.handle.net/1721.1/132617
dc.description.abstractMaterials that sense and respond to biological signals in their environment have a broad range of potential applications in drug delivery, medical devices and diagnostics. Nucleic acids are important biological cues that encode information about organismal identity and clinically relevant phenotypes such as drug resistance. We recently developed a strategy to design nucleic acid–responsive materials using the CRISPR-associated nuclease Cas12a as a user-programmable sensor and material actuator. This approach improves on the sensitivity of current DNA-responsive materials while enabling their rapid repurposing toward new sequence targets. Here, we provide a comprehensive resource for the design, synthesis and actuation of CRISPR-responsive hydrogels. First, we provide guidelines for the synthesis of Cas12a guide RNAs (gRNAs) for in vitro applications. We then outline methods for the synthesis of both polyethylene glycol-DNA (PEG-DNA) and polyacrylamide-DNA (PA-DNA) hydrogels, as well as their controlled degradation using Cas12a for the release of cargos, including small molecules, enzymes, nanoparticles and living cells within hours. Finally, we detail the design and assembly of microfluidic paper-based devices that use Cas12a-sensitive hydrogels to convert DNA inputs into a variety of visual and electronic readouts for use in diagnostics. Following the initial validation of the gRNA and Cas12a components (1 d), the synthesis and testing of either PEG-DNA or PA-DNA hydrogels require 3–4 d of laboratory time. Optional extensions, including the release of primary human cells or the design of the paper-based diagnostic, require an additional 2–3 d each.en_US
dc.description.sponsorshipDefense Threat Reduction Agency (Grant HDTRA1-14-1-0006)en_US
dc.language.isoen
dc.publisherSpringer Science and Business Media LLCen_US
dc.relation.isversionofhttp://dx.doi.org/10.1038/s41596-020-0367-8en_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.sourceProf. Collinsen_US
dc.titleCreating CRISPR-responsive smart materials for diagnostics and programmable cargo releaseen_US
dc.typeArticleen_US
dc.identifier.citationGayet, Raphael V. et al. "Creating CRISPR-responsive smart materials for diagnostics and programmable cargo release." Nature Protocols 15, 9 (August 2020): 3030–3063. © 2020 The Author(s)en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Institute for Medical Engineering & Scienceen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Mechanical Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Synthetic Biology Centeren_US
dc.relation.journalNature Protocolsen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dc.date.updated2021-09-21T13:27:16Z
dspace.orderedauthorsGayet, RV; de Puig, H; English, MA; Soenksen, LR; Nguyen, PQ; Mao, AS; Angenent-Mari, NM; Collins, JJen_US
dspace.date.submission2021-09-21T13:27:18Z
mit.journal.volume15en_US
mit.journal.issue9en_US
mit.licensePUBLISHER_POLICY
mit.metadata.statusCompleteen_US


Files in this item

Thumbnail
Thumbnail

This item appears in the following Collection(s)

Show simple item record