.png){kind=icon}
返回
Creating CRISPR-responsive smart materials for diagnostics and programmable cargo release
.png){kind=icon}
OA
AI
DOI：10.1038/s41596-020-0367-8.png)
.png){kind=icon}
.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
This protocol describes the design and synthesis of CRISPR-responsive smart hydrogels and their actuation for both the controlled release of cargos (small molecules, enzymes, nanoparticles and living cells) and diagnostic applications. Materials 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.
Keyword:
NUCLEIC-ACID DETECTION
HYDROGELS
BIOMATERIALS
EXPRESSION
KINETICS
CAS
AI总结
对已上传原文的论文进行重点信息的提取，主要内容包括：简要概述、研究摘要、背景介绍、关键亮点、图文解析、展望与总结。
期刊
IF:
16
论文数:
4.0K
被引数:
5.6W
机构
引用论文
分享收藏A low-cost paper-based synthetic biology platform for analyzing gut microbiota and host biomarkers.png)
NATURE COMMUNICATIONSOAAI
NATURE COMMUNICATIONSIF15.7
OAAI分享收藏CRISPR-Cas12a has both cis- and trans-cleavage activities on single-stranded DNACRISPR-Cas12a对单链DNA具有顺式和反式切割活性CELL RESEARCHOAAI
CELL RESEARCHIF25.9
OAAI分享收藏Policy, Ethical, Social, and Environmental Considerations of Web3 and the MetaverseWeb3和元宇宙的政策、伦理、社会和环境考虑
OAAI
OAAI分享收藏Real-time observation of DNA target interrogation and product release by the RNA-guided endonuclease CRISPR Cpf1 (Cas12a)通过RNA引导的核酸内切酶CRISPR Cpf1 (Cas12a) 实时观察DNA靶标询问和产物释放OAAI
OAAI分享收藏
查看更多内容