Companion-Probe &amp; Race platform for interrogating nuclear protein and migration of living cells

Sun, Hong; Dong, Zaizai; Zhang, Qingyang; Liu, Bing; Yan, Shi; Wang, Yusen; Yin, Dedong; Ren, Pei‐Gen; Wu, Nan; Chang, Lingqian

doi:10.1016/j.bios.2022.114281

Cited by 8 publications

(2 citation statements)

References 64 publications

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“…In addition, the silver sheet can be bent so that it is in full contact with the solution in the cell culture chamber while facilitating the connection of wires. Generally, the material of the electrode for nanoelectroporation can be ITO-coated glass, − gold, ,,− platinum, , and silver, , which can meet the need for electrical conductivity. The silver sheet was held in place by a clip that was fixed to the iron stand.…”

Section: Methodsmentioning

confidence: 99%

Sensitive Interrogation of Enhancer Activity in Living Cells on a Nanoelectroporation-Probing Platform

et al. 2022

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Enhancers involved in the upregulation of multiple oncogenes play a fundamental role in tumorigenesis and immortalization. Exploring the activity of enhancers in living cells has emerged as a critical path to a deep understanding of cancer properties, further providing important clues to targeted therapy. However, identifying enhancer activity in living cells is challenging due to the double biological barriers of a cell cytoplasmic membrane and a nuclear membrane, limiting the sensitivity and responsiveness of conventional probing methods. In this work, we developed a nanoelectroporation-probing (NP) platform, which enables intranuclear probe delivery for sensitive interrogation of enhancer activity in living cells. The nanoelectroporation biochip achieved highly focused perforation of the cell cytoplasmic membrane and brought about additional driving force to expedite the delivery of probes into the nucleus. The probes targeting enhancer activity (named "PH probe") are programmed with a cyclic amplification strategy and enable an increase in the fluorescence signals over 100-fold within 1 h. The platform was leveraged to detect the activity of CCAT1 enhancers (CCAT1, colon cancer-associated transcript-1, a long noncoding RNA that functions in tumor invasion and metastasis) in cell samples from clinical lung cancer patients, as well as reveal the heterogeneity of enhancers among different patients. The observations may extend the linkages between enhancers and cancer cells while validating the robustness and reliability of the platform for the assay of enhancer activity. This platform will be a promising toolbox with wide applicable potential for the intranuclear study of living cells.

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Section: Methodsmentioning

confidence: 99%

Sensitive Interrogation of Enhancer Activity in Living Cells on a Nanoelectroporation-Probing Platform

et al. 2022

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“…[145][146][147] Additionally, proteins like antibodies, with relatively large hydrodynamic size and specific characteristics, can be delivered through this strategy, [148] along with hydrophilic drugs and more. [19,149,150] For example, the amphiphilic polymer PMAL (poly(maleic anhydride-alt-1-decene) substituted with 3-(dimethylamino) propylamine) creates pores in the cell membrane through electrostatic and hydrophobic interactions with lipids to deliver siRNA. [145] Ishii et al prepared biodegradable polymer microspheres carrying pDNA/liposome complexes.…”

Section: Improving the Delivery Efficiency Of Therapeutic Drugsmentioning

confidence: 99%

Cell Membrane Perforation: Patterns, Mechanisms and Functions

Zhu,

Shi,

Mao

et al. 2024

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Cell membrane is crucial for the cellular activities, and any disruption to it may affect the cells. It is demonstrated that cell membrane perforation is associated with some biological processes like programmed cell death (PCD) and infection of pathogens. Specific developments make it a promising technique to perforate the cell membrane controllably and precisely. The pores on the cell membrane provide direct pathways for the entry and exit of substances, and can also cause cell death, which means reasonable utilization of cell membrane perforation is able to assist intracellular delivery, eliminate diseased or cancerous cells, and bring about other benefits. This review classifies the patterns of cell membrane perforation based on the mechanisms into 1) physical patterns, 2) biological patterns, and 3) chemical patterns, introduces the characterization methods and then summarizes the functions according to the characteristics of reversible and irreversible pores, with the aim of providing a comprehensive summary of the knowledge related to cell membrane perforation and enlightening broad applications in biomedical science.

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Deep learning‐enhanced microwell array biochip for rapid and precise quantification of Cryptococcus subtypes

Tong,

Zeng,

et al. 2024

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Cryptococcus is a family of strongly infectious pathogens that results in a wide variety of symptoms, particularly threatening the patients undergoing the immune‐deficiency or medical treatment. Rapidly identifying Cryptococcus subtypes and accurately quantifying their contents remain urgent needs for infection control and timely therapy. However, traditional detection techniques heavily rely on expensive, specialized instruments, significantly compromising their applicability for large‐scale population screening. In this work, we report a portable microwell array chip platform integrated with a deep learning‐based image recognition program, which enables rapid, precise quantification of the specific subtypes of Cryptococcus. The platform features four zones of microwell arrays preloaded with the subtype‐targeted CRISPR–Cas12a system that avoid dependence on slow, instrumental‐mediated target amplification, achieving rapid (10 min), high specificity for identifying the sequence of Cryptococcus. The deep learning‐based image recognition program utilizing segment anything model (SAM) significantly enhances automation and accuracy in identifying target concentrations, which eventually achieves ultra‐low limit of detection (0.5 pM) by personal smartphones. This platform can be further customized to adapt to various scenarios in clinical settings.

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Companion-Probe & Race platform for interrogating nuclear protein and migration of living cells

Cited by 8 publications

References 64 publications

Sensitive Interrogation of Enhancer Activity in Living Cells on a Nanoelectroporation-Probing Platform

Sensitive Interrogation of Enhancer Activity in Living Cells on a Nanoelectroporation-Probing Platform

Cell Membrane Perforation: Patterns, Mechanisms and Functions

Deep learning‐enhanced microwell array biochip for rapid and precise quantification of Cryptococcus subtypes

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