Epigenetics is one of the many ways in which cells regulate gene expression, and it studies the mechanisms by which genes are switched on or off without changing their genetic code or DNA sequence. The regulation of gene transcription level plays a vital role in the process of gene expression regulation and is a key factor affecting the level of gene expression. Therefore, the study of gene transcription level regulation has always been a hot topic in gene expression regulation research. Lifeasible can provide you with various technologies based on methylated DNA immunoprecipitation (MeDIP) and gene transcriptional activity analysis services so that you can detect the expression of key genes and obtain the methylation status of genes.

Real-time fluorescence quantitative PCR technology combined with MeDIP (MeDIP-RT-qPCR)

RT-qPCR is a method of adding fluorescent reagents into the PCR reaction system, detecting the PCR process in real time using fluorescent signals, and finally quantify the unknown templates by standard curves. RT-qPCR can quantify the concentration of the starting templates in a specific, sensitive, rapid, and highly reproducible way. Based on the characteristics of RT-qPCR technology, we combine it with MeDIP technology, first, enrich methylated DNA fragments by MeDIP technology, then detect them by RT-qPCR technology, and then obtain the gene expression data of the specific locus and the precise value of methylation level in the detected biological samples after data analysis and processing. The method we provide has high specificity, does not require labeled probes, and has high accuracy, good reproducibility, and high sensitivity.

Microarray technology combined with MeDIP (MeDIP-chip)

We use MeDIP to enrich methylated DNA fragments, which are labeled separately from whole genome controls and then mixed and hybridized to the designed microarrays, and the hybridization signals are detected by a high-resolution microarray scanner. The hybridization results are then subjected to data extraction, normalization, peak analysis, and reporting. This helps you to obtain gene expression data as well as methylation data.

Analysis of transcriptional activity of genes

Studying the transcriptional regulation of a gene generally involves studying the activity of the gene's promoter, the activity of the transcription factors that regulate the gene, and the activity of the transcriptional co-regulators involved in the regulation. When we examine the regulation of a promoter segment by a transcription factor, we will co-transfect cells with a reporter gene vector containing the regulation of the promoter and the transcription factor and evaluate the function of the transcription factor by detecting the level of the reporter gene. This will both qualitatively reflect whether the transcription factor regulates that promoter and quantitatively assess how active the transcription factor is. If a transcription co-regulatory factor is co-transfected with the promoter, we can also determine the nature and activity of the transcription co-regulatory factor by detecting the level of the reporter gene.

Flow of activity analysis of gene promoters, activity analysis of transcription factors, and activity analysis of transcriptional co-regulatory factors:

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