Various Strategies for Improved Signal-to-Noise Ratio in CRISPR-Based Live Cell Imaging

Abstract

The various behaviors of cells are closely related to the temporal and spatial dynamics of chromatin. Live cell imaging techniques for chromatin can provide insight into gene expression at the three dimension of a nucleus and offer the novel knowledge in gene regulation for medical diagnosis, crop improvement, and environmental remediation. However, traditional chromatin imaging techniques that rely on the insertion of a target gene are unable to directly analyze the chromatin dynamics in living cell nuclei. CRISPR-based live imaging, with its high accuracy, speed, and low perturbation, has emerged as a reliable tool for high-resolution imaging of the specific chromatin in the nucleus. This review paper introduces the benefits of CRISPR-based live imaging for chromatin dynamics, classifies and analyzes various successful schemes used to improve the signal-to-noise ratio of this method, and discusses future trends in the field.