• Diatrack particle tracking software: Review of applications and performance evaluation

      Vallotton, Pascal; van Oijen, Antoine M.; Whitchurch, Cynthia B.; Gelfand, Vladimir; Yeo, Leslie; Tsiavaliaris, Georgios; Heinrich, Stephanie; Dultz, Elisa; Weis, Karsten; Grunwald, David (2017-12-01)
      Object tracking is an instrumental tool supporting studies of cellular trafficking. There are three challenges in object tracking: the identification of targets; the precise determination of their position and boundaries; and the assembly of correct trajectories. This last challenge is particularly relevant when dealing with densely populated images with low signal-to-noise ratios-conditions that are often encountered in applications such as organelle tracking, virus particle tracking or single-molecule imaging. We have developed a set of methods that can handle a wide variety of signal complexities. They are compiled into a free software package called Diatrack. Here we review its main features and utility in a range of applications, providing a survey of the dynamic imaging field together with recommendations for effective use. The performance of our framework is shown to compare favorably to a wide selection of custom-developed algorithms, whether in terms of localization precision, processing speed or correctness of tracks.
    • High-Resolution Mapping of Multiway Enhancer-Promoter Interactions Regulating Pathogen Detection

      Vangala, Pranitha; Murphy, Rachel; Quinodoz, Sofia A.; Gellatly, Kyle J.; McDonel, Patrick E.; Guttman, Mitchell; Garber, Manuel (2020-10-15)
      Eukaryotic gene expression regulation involves thousands of distal regulatory elements. Understanding the quantitative contribution of individual enhancers to gene expression is critical for assessing the role of disease-associated genetic risk variants. Yet, we lack the ability to accurately link genes with their distal regulatory elements. To address this, we used 3D enhancer-promoter (E-P) associations identified using split-pool recognition of interactions by tag extension (SPRITE) to build a predictive model of gene expression. Our model dramatically outperforms models using genomic proximity and can be used to determine the quantitative impact of enhancer loss on gene expression in different genetic backgrounds. We show that genes that form stable E-P hubs have less cell-to-cell variability in gene expression. Finally, we identified transcription factors that regulate stimulation-dependent E-P interactions. Together, our results provide a framework for understanding quantitative contributions of E-P interactions and associated genetic variants to gene expression.