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Updated: Jun 23, 2025

Quantitative Fluorescence In Situ Hybridization FISH and Immunofluorescence IF of Specific Gene Products in KSHV-Infected Cells

Quantitative Fluorescence In Situ Hybridization FISH and Immunofluorescence IF of Specific Gene Products in KSHV-Infected Cells

Published on: August 27, 2019

卡波西肉瘤相关的疹病毒转录组的高密度分辨率识别了通过长距离转录和替代拼接生成的新型转录异型

Ritu Shekhar¹, Tina O'Grady², Netanya Keil^1,3

¹Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL, USA.

Nucleic acids research

|June 26, 2024

概括

研究人员开发了一种新方法来分析卡波西的瘤相关疹病毒 (KSHV) 转录,揭示了994个独特的KSHV转录和新的基因表达见解.

科学领域:

病毒学病毒学
基因组学就是基因组学.
文字转录学 (Transcriptomics) 是一个学科.

背景情况:

卡波西肉瘤相关的疹病毒 (KSHV) 导致卡波西肉瘤和B细胞恶性瘤.
人类疹病毒的高分辨率转录组揭示了密集的基因编码.
在性复制过程中,KSHV转录分析受到低转录百分比的限制.

研究的目的:

开发一种目标丰富方法来增加KSHV特定的测序读数.
通过集成多平台数据 (TRIMD) 管道将丰富与转录组分辨率结合起来,用于转录注释.
为了生成第一个高密度的KSHV光学转录组.

主要方法:

开发了针对短读和长读测序平台的目标丰富方法.
与TRIMD管道进行集成丰富,用于转录结构注释.
TRIMD管道使用长读测序架,由Illumina RNA-Seq和deepCAGE数据验证.

主要成果:

确定了994个独特的KSHV转录,创建了第一个高密度的KSHV溶性转录组.
发现了许多新的KSHV转录异型,包括替代的UTR,拼接连接和ORF.
确定了转录起始点,可能会增加与延迟相关的基因表达.

结论:

这项研究为KSHV基因表达调节提供了前所未有的见解.
新的KSHV转录异型提供了对病毒基因表达的更深入的理解.
这些发现可能会揭示卡波西肉瘤瘤中延迟相关基因的表达.

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Detection of the Genome and Transcripts of a Persistent DNA Virus in Neuronal Tissues by Fluorescent In situ Hybridization Combined with Immunostaining

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