相关实验视频

Updated: May 31, 2025

Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity

Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity

Published on: May 1, 2018

dsDAP:一种高效的方法,在哺乳动物细胞中构建高丰度DNA编码的库

Kaili Zhang¹, Yi Wang¹, Shuze Jiang¹

¹Department of Molecular Pharmacology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, Tianjin's Clinical Research Center for Cancer, Tianjin 300060, China.

International journal of biological macromolecules

|January 22, 2025

概括

我们开发了一种新的DNA组装方法,即dsDNA-assembly-PCR (dsDAP),用于在哺乳动物细胞中创建高丰度的DNA编码库. 这种方法提高了功能基因组学和药物发现的灵活性和效率.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
生物技术是生物技术.

背景情况:

用DNA编码的库对于高通量查和功能基因组学是必不可少的.
在哺乳动物细胞中构建高丰度图书馆面临重大挑战.

研究的目的:

引入dsDNA组合-PCR (dsDAP),一种新的吉布森组合-PCR策略,用于在哺乳动物细胞中高效地构建DNA编码库.
调查翻译启动序列 (TIS) 对蛋白质表达的影响,并探索图书馆稀释对基因型-表型相关性的影响.

主要方法:

开发并应用了dSDNA组装PCR (dsDAP) 策略,用于DNA编码库的创建.
利用CRISPR-Cas9和piggyBac系统在HEK293T细胞中进行基因组整合.
分析了TIS核酸和库稀释对蛋白质表达和基因型-表型相关性的影响.

主要成果:

与现有方法相比,dSDAP的灵活性和效率得到了提高.
证实了TIS中特定核酸的关键作用,特别是氨酸在-3位置,用于高蛋白表达.
展示了 dsDAP 在哺乳动物系统中的高丰度库构建的实用性.

结论:

dsDAP策略提供了一种多功能和高效的解决方案,用于在哺乳动物细胞中构建高丰度的DNA编码库,克服了以前技术的局限性.
这种方法对功能基因组学,药物发现和基因调节研究有重大影响.

关键词:

这就是CRISPR-Cas9的特征.一个DNA编码的图书馆.大规模并行测试 (MPAs)大规模并行报告测试 (MPRAs)翻译启动序列 (TISs) 是指翻译开始的序列.dsDNA组合-PCR (dsDAP) 是一个很好的方法.在 PiggyBac 里面,你会看到一个猪.

更多相关视频

Author Spotlight: Simplifying Genome-Wide Plasmid Library Construction Using CRISPRmass

Author Spotlight: Simplifying Genome-Wide Plasmid Library Construction Using CRISPRmass

Published on: May 17, 2024

Author Spotlight: An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations

Author Spotlight: An Integrated Workflow to Study the Promoter-Centric Spatio-Temporal Genome Architecture in Scarce Cell Populations

Published on: April 21, 2023

相关概念视频

DNA Isolation

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...