No abstract

Genome architecture and organization play critical roles in cell life. However, it remains largely unknown how genomic loci are dynamically coordinated to regulate gene expression and determine cell fate at the single cell level. We have developed an inducible system which allows Simultaneous Imaging and Manipulation of genomic loci by Biomolecular Assemblies (SIMBA) in living cells. In SIMBA, the human heterochromatin protein 1α (HP1α) is fused to mCherry and FRB, which can be induced to form biomolecular assemblies (BAs) with FKBP-scFv, guided to specific genomic loci by a nuclease-defective Cas9 (dCas9) or a transcriptional factor (TF) carrying tandem repeats of SunTag. The induced BAs can not only enhance the imaging signals at target genomic loci using a single sgRNA, either at repetitive or non-repetitive sequences, but also recruit epigenetic modulators such as histone methyltransferase SUV39H1 to locally repress transcription. As such, SIMBA can be applied to simultaneously visualize and manipulate, in principle, any genomic locus with controllable timing in living cells.

From basic studies in understanding the role of signaling pathways to therapeutic applications in engineering new cellular functions, efficient and safe techniques to monitor and modulate molecular targets from cells to organs have been extensively developed. The developmental advancement of engineering devices such as microscope and ultrasonic transducers allows us to investigate biological processes at different scales. Synthetic biology has further emerged recently as a powerful platform for the development of new diagnostic and therapeutic molecular tools. The synergetic amalgamation between engineering tools and synthetic biology has rapidly become a new front in the field of bioengineering and biotechnology. In this review, ultrasound and its generated mechanical perturbation are introduced to serve as a non-invasive engineering approach and, integrated with synthetic biology, to remotely control signaling and genetic activities for the guidance of cellular functions deep inside tissue with high spatiotemporal resolutions. This ultrasound-based approach together with synthetic biology has been applied in immunotherapy, neuroscience, and gene delivery, paving the way for the development of next-generation therapeutic tools.

Despite its success in treating hematologic malignancies, chimeric antigen receptor (CAR) T cell therapy faces two major challenges which hinder its broader applications: the limited effectiveness against solid tumors and the nonspecific toxicities. To address these concerns, researchers have used synthetic biology approaches to develop optimization strategies. In this review, we discuss recent improvements on the CAR and other non-CAR molecules aimed to enhance CAR T cell efficacy and safety. We also highlight the development of different types of inducible CAR T cells that can be controlled by environmental cues and/or external stimuli. These advancements are bringing CAR T therapy one step closer to safer and wider applications, especially for solid tumors.