Pub Date : 2024-03-07DOI: 10.1109/TNB.2024.3373597
Jiayu Sun;Yang Jiao;Fei Pan;Shuk Han Cheng;Dong Sun
The efficient application of the newly developed gene-editing method CRISPR/Cas9 requires more accurate intracellular gene delivery. Traditional delivery approaches, such as lipotransfection and non-viral delivery methods, must contend with major problems to overcome the drawbacks of low efficiency, high toxicity, and cell-type dependency. The high-throughput microdroplet-based single-cell transfection method presented herein provides an alternative method for delivering genome-editing reagents into single living cells. By accurately controlling the number of exogenous plasmids in microdroplets, this method can achieve high-efficiency delivery of nucleic acids to different types of single cells. This paper presents a high-throughput quantitative DNA transfection method for single cells and explores the optimal DNA transfection conditions for specific cell lines. The transfection efficiency of cells at different concentrations of DNA in microdroplets is measured. Under the optimized transfection conditions, the method is used to construct gene-knockout cancer cell lines to determine specific gene functions through the CRISPR/Cas9 knockout system. In a case study, the migration ability of TRIM72 knockout cancer cells is inhibited, and the tumorigenicity of cells in a zebrafish tumor model is reduced. A single-cell microfluidic chip is designed to achieve CRISPR/Cas9 DNA transfection, dramatically improving the transfection efficiency of difficult-to-transfect cells. This research demonstrates that the microdroplet method developed herein has a unique advantage in CRISPR/Cas9 gene-editing applications.
新开发的基因编辑方法 CRISPR/Cas9 的高效应用需要更精确的细胞内基因递送。传统的递送方法,如脂质转染和非病毒递送方法,必须克服低效率、高毒性和细胞类型依赖性等主要问题。本文介绍的基于微滴的高通量单细胞转染方法为向单个活细胞递送基因组编辑试剂提供了另一种方法。通过精确控制微滴中外源质粒的数量,这种方法可以实现向不同类型的单细胞高效递送核酸。本文介绍了一种高通量定量 DNA 单细胞转染方法,并探讨了特定细胞系的最佳 DNA 转染条件。测量了微滴中不同浓度 DNA 对细胞的转染效率。在优化的转染条件下,该方法被用于构建基因敲除癌症细胞系,通过 CRISPR/Cas9 基因敲除系统确定特定基因的功能。在一项案例研究中,TRIM72基因敲除癌细胞的迁移能力受到抑制,斑马鱼肿瘤模型中细胞的致瘤性降低。设计了一种单细胞微流控芯片来实现 CRISPR/Cas9 DNA 转染,大大提高了难以转染细胞的转染效率。这项研究表明,本文开发的微滴方法在 CRISPR/Cas9 基因编辑应用中具有独特的优势。
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Pub Date : 2024-03-06DOI: 10.1109/TNB.2024.3396142
Zhao-Cai Wang;Xian Wu;Kun Liang;Tun-Hua Wu
The traveling car renter problem (TCRP) is a variant of the Traveling Salesman Problem (TSP) wherein the salesman utilizes rented cars for travel. The primary objective of this problem is to identify a solution that minimizes the cumulative operating costs. Given its classification as a non-deterministic polynomial (NP) problem, traditional computers are not proficient in effectively resolving it. Conversely, DNA computing exhibits unparalleled advantages when confronted with NP-hard problems. This paper presents a DNA algorithm, based on the Adleman-Lipton model, as a proposed approach to address TCRP. The solution for TCRP can be acquired by following a series of fundamental steps, including coding, interaction, and extraction. The time computing complexity of the proposed DNA algorithm is $boldsymbol {O}(boldsymbol {n}^{boldsymbol {2}}boldsymbol {m})$