被遗忘的变量? 安乐死方法和样本储存条件是否会影响生物体的转录组 - - 在猪的多个组织上对基因表达的分析
B Chakkingal Bhaskaran1, R Meyermans2, W Gorssen2
1Department of Biosystems, Centre for Animal Breeding and Genetics, KU Leuven, Kasteelpark Arenberg 30, Box 2472, Leuven, 3001, Belgium. bimalcb98@gmail.com.
BMC genomics
|December 13, 2023
概括
气安乐死 (ANOXIA) 是转录基因研究中对小猪的合适替代方案. 然而,安乐死方法和组织存储条件都可能影响基因表达,需要仔细考虑,以避免误解结果.
科学领域:
- 动物研究方法论动物研究方法论
- 分子生物学分子生物学
- 文字转录学 (Transcriptomics) 是一个学科.
背景情况:
- 转录组研究需要在安乐死后采集新鲜组织.
- 安乐死方法可以引入意想不到的基因表达变异.
- 调查安乐死和储存对基因表达的影响对于可靠的转录基因数据至关重要.
研究的目的:
- 为了比较气 (ANOXIA) 与T-61® (T61) 安乐死方法,用于小猪转录基因研究.
- 分析RNAlaterTM (RL) 和液 (LN2) 快速结对基因表达特征的影响.
- 在转录组学研究中识别生物变异的潜在来源.
主要方法:
- 3'mRNA-Seq对脑下垂体,下丘脑,肝脏和肺组织的数据分析.
- 不同安乐死方法之间的基因表达特征的比较.
- 在两个常见的组织存储条件下 (RL和LN2) 评估RNA完整性和基因表达.
主要成果:
- 在ANOXIA和T61安乐死方法之间没有观察到蛋白质编码基因的显著差异表达.
- 在ANOXIA组的垂体样本中检测到小核RNAs (snRNAs) 的水平升高.
- RNAlaterTM (RL) 提供了优越的RNA保存 (更高的RIN值),但影响基因表达不同于液 (LN2) 快速结.
结论:
- 氧无氧是一种可行的安乐死方法,用于小猪转录学研究,对蛋白质编码基因的影响最小.
- 非蛋白质编码RNAs,如snRNAs,可能会受到安乐死方法的影响,可能会影响下游基因表达.
- 安乐死和组织存储方法都代表了重要的生物变异来源,必须考虑这些变化,以防止对转录基因数据的误解.
相关概念视频
02:51mRNA Stability and Gene Expression
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Cis-acting Elements involved in mRNA stability
01:36What is Gene Expression?
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then processed and...
01:30General Transcription Factors
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
02:51mRNA Stability and Gene Expression
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Cis-acting Elements involved in mRNA stability