Massively Parallel Reporter Assays for High-Throughput In Vivo Analysis of Cis-Regulatory Elements.
Yanjiang ZhengNathan J VanDusenPublished in: Journal of cardiovascular development and disease (2023)
The rapid improvement of descriptive genomic technologies has fueled a dramatic increase in hypothesized connections between cardiovascular gene expression and phenotypes. However, in vivo testing of these hypotheses has predominantly been relegated to slow, expensive, and linear generation of genetically modified mice. In the study of genomic cis -regulatory elements, generation of mice featuring transgenic reporters or cis -regulatory element knockout remains the standard approach. While the data obtained is of high quality, the approach is insufficient to keep pace with candidate identification and therefore results in biases introduced during the selection of candidates for validation. However, recent advances across a range of disciplines are converging to enable functional genomic assays that can be conducted in a high-throughput manner. Here, we review one such method, massively parallel reporter assays (MPRAs), in which the activities of thousands of candidate genomic regulatory elements are simultaneously assessed via the next-generation sequencing of a barcoded reporter transcript. We discuss best practices for MPRA design and use, with a focus on practical considerations, and review how this emerging technology has been successfully deployed in vivo. Finally, we discuss how MPRAs are likely to evolve and be used in future cardiovascular research.
Keyphrases
- high throughput
- copy number
- gene expression
- transcription factor
- crispr cas
- single cell
- primary care
- rna seq
- high fat diet induced
- type diabetes
- wild type
- skeletal muscle
- current status
- cross sectional
- big data
- adipose tissue
- electronic health record
- machine learning
- african american
- deep learning
- sensitive detection
- clinical evaluation
- circulating tumor cells