Gene Expression Profiling-Based Screening

Transcriptional or expression profiling of intact and isolated tissues is an essential part of the drug development process. This has significant implications for target identification, validation, compound selection, pharmacogenomics, biomarker development, clinical trial evaluation, and toxicology. Ace Therapeutics has established a complete gene expression profiling analysis platform to identify and quantify ocular gene expression levels, and help customers identify new drug targets through systematic analysis of genome-wide expression profiles and differential gene expression profiles.

Gene Expression Profiling-Based Target Screening

Gene Expression Profiling Introduction

Gene expression profiling is a method in the field of molecular biology that measures gene expression in cells using cDNA, expressed sequence tags, or oligonucleotide microarrays. It is able to measure a large number of genes at once to build an overall view of cell function. From this, it can distinguish between dividing cells and those responding to signature treatments. Gene expression profiling also helps in understanding eye disease pathogenesis, physiological responses to drugs, and therapeutic effects. For target identification purposes, compare gene expression profiles between drug-treated cells and no-drug control cells. Different expression patterns provide scientists with valuable insights to discover drug targets, elucidate the molecular basis of drug-associated phenotypic changes, and select appropriate targets for in-depth study.

Fig. 1. A diagrammatic overview of the considerations for designing a successful RNA-seq experiment for differential gene expression analysis.(Owen N, et al., 2019)

Services Overview

Ace Therapeutics has a high-quality team that has rich experience in many fields such as ophthalmology, genetics, molecular biology, and biological informatics. Our researchers can perform genetic expression analysis at any of several different levels of gene expression: transcriptional, post-transcriptional, translational, and post-translational protein modification. A variety of molecular biology technology and experimental methods, including the Northern Miracle, DNA micro-array, real-time polymerase chain reaction, chromatin immune precipitation (Chip), Western blotting, mass spectrometry and immunization measurement, are adopted to support our global customers' projects.

Our Capabilities

At Ace Therapeutics, our gene expression analysis covers multiple fields such as RNA expression and promoter analysis, and provides a series of corresponding research methods. For different eye conditions and diseases, we develop customized gene expression profiling services

RNA Expression Analysis

Northern blotting, DNA microarray and real-time polymerase chain reaction (RT-PCR) are our main tools for RNA expression analysis.

Northern Blot Analysis

Northern blot analysis, also known as Northern Blot analysis, is a laboratory analysis method used to study RNA. This tag allows any RNA fragment that contains a sequence complementary to the DNA probe sequence to be visualized on a northern blot. Ace Therapeutics uses northern blot analysis to identify whether genes are overexpressed or underexpressed in eye cancer cells, compared to normal cells. We provide one-stop northern blot analysis services, including extraction of total RNA from homogenized tissue samples or cells, separation of eukaryotic mRNA, and gel electrophoresis analysis.

DNA Microarray Analysis

As an exciting tool for gene expression profiling, DNA microarrays are a powerful tool for RNA expression analysis at Ace Therapeutics. In this method, your specifically labeled RNA-expressing sample is hybridized with immobilized DNA probes to measure gene expression levels. We offer a variety of methods for labeling cDNA or cRNA, including: incorporation of a fluorescently marked nucleotide during synthesis, incorporation of a biotin-tagged nucleotide. In addition, our geneticists also use microarrays in conjunction with chromatin immunosuppression to identify transcription factor binding sites.

RT-PCR Analysis

PCR represents the method of choice for analyzing gene expression in moderate numbers of genes ranging from a few to thousands of samples. Ace Therapeutics uses digital PCR, real-time PCR and quantitative PCR for ocular gene expression profiling. We offer RT-PCR services to verify gene expression patterns. Raw data obtained from RT-PCR were converted into an informative format for better understanding.

RNA Sequencing Analysis

High-throughput RNA sequencing (RNA-Seq) sequencing technology is capable of capturing many sub-types of RNA molecules and has become the gold standard for transcriptome gene expression quantification. Our geneticists use RNA-Seq technology to characterize all transnational activity (coding and non-coding) associated with ocular disease, including low expressed genes and novel transcripts. Model systems, such as animal or cell-based systems, provide a valuable resource for research around the globe. We provide precise measurements of strand orientation, uniform coverage, and high-confidence mapping of alternative transcripts and gene fusions. It offers a promising way to select targets of interest.

Services Process

Fig.2. Workflow for gene expression profiling.

Sample Preparation: Obtain the required ocular tissues from the oculars of the experimental subjects (rats, mice), including conjunctival cells, retinal tissues, corneas, conjunctivas, and so on.
RNA Extraction: Extract total RNA from samples such as conjunctival cells, retinal tissues, corneas and conjunctivas.
RNA Sample Evaluation: RNA yield and purity were evaluated using a NanoDrop ND-100 spectrophotometer.
Reverse Transcription Analysis: The use of multiple technologies for RNA expression profiling, along with the use of RNA-Seq technology, will yield a comprehensive understanding of all transnational activity (coding and non-coding) associated with ocular diseases.
Bioinformatics: Analysis of gene expression differences between responder and non-responder groups.
Gene Expression Profiling: Identify potential drug targets for different eye diseases based on differentially expressed genes.

Thanks to world-class equipment and advanced technology platforms, Ace Therapeutics provides global customers with one-stop ocular gene expression profiling services to identify new drug targets. In addition, our team of experienced geneticists will help you choose the most suitable solution based on your project needs. If you are interested in our services or want to know more information, please feel free to contact us. We look forward to working with you.

Reference

Owen N, Moosajee M. RNA-sequencing in ophthalmology research: considerations for experimental design and analysis. Ther Adv Ophthalmol. 2019:15;11:2515841419835460.

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