Using Droplet Digital PCR to Analyze Allele-Specific RNA Expression

Area of Science:

• Genetics
• Molecular Biology
• Bioinformatics

Background:

• Genome-wide association studies (GWAS) identify common alleles linked to human traits and diseases.
• Many disease-associated variants reside in regulatory regions, influencing gene expression.
• Understanding allele-specific expression is crucial for interpreting genetic associations.

Purpose of the Study:

• To present a protocol for precisely measuring allele-specific expression (ASE) of individual genes.
• To enable the quantification of allelic skew, reflecting differential expression between alleles.
• To provide a well-controlled method for identifying allelic influences on gene expression.

Main Methods:

• Targeting nucleotide differences between alleles within an individual's diploid genome.
• Developing and optimizing assays for precise measurement of allele expression.
• Analyzing data to determine the extent of allelic skew.

Main Results:

• A robust protocol for measuring allele-specific expression is described.
• The method allows for the quantification of allelic skew.
• Guidance is provided on assay design, reaction optimization, and data interpretation.

Conclusions:

• This protocol offers a precise approach to study allele-specific expression.
• It facilitates the investigation of how genetic variants impact gene expression and phenotypes.
• The method is valuable for research in human genetics and disease association studies.