先进分子生物学技术的实用方法。

4区 生物学 Q4 Biochemistry, Genetics and Molecular Biology
Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2025-03-13 DOI:10.1016/bs.mcb.2025.02.022
Yakhlesh Gupta, Kunzang Chosdol
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引用次数: 0

摘要

近年来,随着强大的技术的发展,分子生物学领域取得了巨大的进步,可以在分子水平上深入探索细胞过程。本章“高级分子生物学技术”提供了分子技术的详细方案。我们从CRISPR-Cas9基因组编辑开始,这是一种精确有效的基因操作变革工具,可以在各种生物体中实现靶向突变和基因敲除。然后讨论了通过Real-Time PCR进行基因扩增,强调了其量化基因表达和高灵敏度检测罕见遗传变异的能力。流式细胞术紧随其后,为分析基于特定标记的细胞群体提供了一个强大的平台,使免疫细胞、癌症诊断和细胞周期分析的研究成为可能。染色质免疫沉淀测序(ChIP-Seq)被探索作为一种绘制蛋白质- dna相互作用的方法,为基因调控和表观遗传修饰提供见解。本章还涵盖了单细胞RNA测序(scRNA-Seq),这是一种在单细胞水平上分析基因表达的突破性技术,允许发现细胞异质性和复杂的生物过程。接下来,我们通过基于质谱的分析探索蛋白质组学,通过鉴定和定量复杂样品中的蛋白质,提供详细的蛋白质组学表征和生物标志物发现。最后,荧光原位杂交(FISH)是一种可视化的完整细胞或组织内特定核酸序列的空间定位的方法。总之,这些先进的分子生物学技术为健康、疾病和细胞功能的分子机制提供了无与伦比的精确度和洞察力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Practical approaches to advanced molecular biology techniques.

The field of molecular biology has undergone tremendous advancements in recent years, with the development of powerful techniques that allow for in-depth exploration of cellular processes at the molecular level. This chapter, "Advanced Molecular Biology Techniques," provides a detailed protocol of the molecular techniques. We begin with CRISPR-Cas9 genome editing, a transformative tool for precise and efficient gene manipulation, enabling targeted mutations and gene knockouts in various organisms. Gene amplification via Real-Time PCR is then discussed, highlighting its ability to quantify gene expression and detect rare genetic variants with high sensitivity. Flowcytometry follows, offering a robust platform for analyzing cellular populations based on specific markers, enabling the study of immune cells, cancer diagnostics, and cell cycle analysis. Chromatin Immunoprecipitation Sequencing (ChIP-Seq) is explored as a method for mapping protein-DNA interactions, providing insights into gene regulation and epigenetic modifications. The chapter also covers Single-cell RNA sequencing (scRNA-Seq), a groundbreaking technique for profiling gene expression at the single-cell level, allowing for the discovery of cell heterogeneity and complex biological processes. Next, we explore into proteomics through Mass Spectrometry-Based Analysis, which offers detailed proteome characterization and biomarker discovery by identifying and quantifying proteins in complex samples. Finally, Fluorescence In Situ Hybridization (FISH) is discussed as a method for visualizing the spatial localization of specific nucleic acid sequences within intact cells or tissues. Together, these advanced molecular biology techniques offer unparalleled precision and insight into the molecular mechanisms underlying health, disease, and cellular function.

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来源期刊
Methods in cell biology
Methods in cell biology 生物-细胞生物学
CiteScore
3.10
自引率
0.00%
发文量
125
审稿时长
3 months
期刊介绍: For over fifty years, Methods in Cell Biology has helped researchers answer the question "What method should I use to study this cell biology problem?" Edited by leaders in the field, each thematic volume provides proven, state-of-art techniques, along with relevant historical background and theory, to aid researchers in efficient design and effective implementation of experimental methodologies. Over its many years of publication, Methods in Cell Biology has built up a deep library of biological methods to study model developmental organisms, organelles and cell systems, as well as comprehensive coverage of microscopy and other analytical approaches.
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