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单分子光现场杂交 (smFISH) 用于检测病原体Candida albicans中的RNA

Sander van Otterdijk¹, Maryam Motealleh¹, Zixu Wang¹

¹Systems Biology Lab, A-LIFE department, Amsterdam Institute of Molecular and Life Sciences (AIMMS), Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.

Methods in molecular biology (Clifton, N.J.)

|March 19, 2024

概括

这项研究引入了一种单分子RNA成像协议,以精确测量单个Candida albicans细胞中的mRNA水平. 这种方法有助于理解真菌形态变化和感染的分子基础.

科学领域:

微生物学微生物学
分子生物学分子生物学
菌类学菌类学是指菌类学.

背景情况:

菌 (Candida albicans) 是人类主要的真菌病原体.
它的致病性与可逆形态变化 (酵母菌,伪菌,菌) 有关.
了解这些开关的分子调节对于对抗感染至关重要.

研究的目的:

开发和详细介绍一个单细胞,单分子RNA成像协议,用于Candida albicans.
为了在完整的,形态上不同的细胞中实现绝对mRNA量化.
为了促进在形态转变期间基因表达的研究.

主要方法:

优化单分子光在位杂交 (smFISH) 对于C. albicans.
详细的协议步骤:细胞生长,固定,杂交,成像,细分和mRNA点位分析.
在不同细胞类型和条件下对特定mRNA (CLB2,EFG1) 的量化.

主要成果:

smFISH协议允许在单个C. albicans细胞中准确量化mRNA.
在酵母,伪基因和形之间观察到不同的基因表达.
成功测量了CLB2和EFG1mRNAs的表达变化.

结论:

这种单细胞RNA成像方法为C. albicans的分子表征提供了强大的工具.
该协议可用于研究其他线状真菌,包括致病性和非致病性真菌.
它提供了一条更好地理解和潜在地针对C. albicans感染的途径.

关键词:

在Candida albicans中使用.菌类病原体的真菌病原体 RNA局部化的RNA局部化一个单细胞成像技术.单分子RNA FISH 的研究结果.偷偷捕捞的鱼类是什么意思

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相关概念视频

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

In-situ Hybridization

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...