在骨质基因分化下,RPLP0/TBP是人类牙髓干细胞最稳定的参考基因
Daniel B Ferreira1, Leticia M Gasparoni1, Cristiane F Bronzeri1
1Laboratory of Extracellular Matrix Biology and Cellular Interaction, Department of Anatomy, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-000, São Paulo, Brazil.
World journal of stem cells
|July 1, 2024
概括
RPLP0和TBP是人类牙髓干细胞骨质分化的最稳定的参考基因,而TUB和B2M是不稳定的. 这一发现确保了组织工程中精确的基因表达分析.
科学领域:
- 干细胞生物学 干细胞生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 在定量实时聚合酶链反应 (RT-qPCR) 中精确的基因表达正常化依赖于稳定的参考基因 (RGs).
- 许多研究在没有验证的情况下错误地使用常见的RGs (例如GAPDH,18SrRNA,β-actin),从而损害了数据的可信性.
- 组织工程需要精确的基因表达数据来制定有效的治疗策略.
研究的目的:
- 在人类牙髓干细胞 (DPSCs) 的骨质基因分化过程中确定RT-qPCR最稳定的参考基因.
- 为经历骨质生成的DPSC提供可靠的基因表达正常化标准.
- 增强基于RT-qPCR的牙组织工程研究的可信度.
主要方法:
- 人类DPSC在不分化和骨质分化条件下培养了35天.
- 10个候选参考基因的基因表达用RT-qPCR在7天间隔进行了评估.
- 数据使用多个算法 (ΔCt,geNorm,NormFinder,BestKeeper) 进行分析,并通过RefFinder进行排名.
主要成果:
- 核糖体蛋白,大,P0 (RPLP0) 和TATA结合蛋白 (TBP) 被确定为最稳定的参考基因.
- 图布林 (TUB) 和β-2-微球蛋白 (B2M) 被发现是最不稳定的参考基因.
- RPLP0/TBP在各种分析算法和子组中表现出一致的稳定性.
结论:
- RPLP0和TBP被证实是单层培养中人类DPSC的长期骨质基因分化最稳定的参考基因.
- TUB和B2M被认为是不稳定的,应该避免在这种情况下进行正常化.
- 这项研究建立了可靠的基因选择标准,用于在牙髓干细胞研究中准确的基因表达研究.
相关概念视频
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...
Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Multi-species Conserved Sequences
Next-generation sequencing technologies have created large genomic databases of a variety of animals and plants. Ever since the human genome project was completed, scientists studied the genome of primates, mammals, and other phylogenetically distant living beings. Such large-scale studies have provided new insights into the evolutionary relationship between organisms.
Although the genome of each species varies greatly from each other, a few sequences are highly conserved. Such conserved DNA...
Although the genome of each species varies greatly from each other, a few sequences are highly conserved. Such conserved DNA...
General 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...


