ACS synthetic biology [ACS Synth Biol] 2021 Jun 18; Vol. 10 (6), pp. 1300-1307. Date of Electronic Publication: 2021 May 20.
Cyanobacteria are promising microbial hosts for the production of diverse biofuels and biochemicals. However, compared to other model microbial hosts such as Escherichia coli and yeast, it takes a long time to genetically modify cyanobacteria. One way to efficiently engineer cyanobacteria while minimizing genetic engineering would be to develop a fast, high-throughput prototyping tool for cyanobacteria. In this study, we developed a CRISPR/Cas12a-based assay coupled with cyanobacteria cell-free systems to rapidly prototype promoter characteristics. Using this newly developed assay, we demonstrated cyanobacteria cell-free transcription for the first time and confirmed a positive correlation between the in vitro and in vivo transcription performance. Furthermore, we generated a synthetic promoter library and evaluated the characteristics of promoter subregions by using the assay. Varied promoter strength derived from random mutations were rapidly and effectively measured in a high-throughput way. We believe that this study offers an easily applicable and rapid prototyping platform to characterize promoters for cyanobacterial engineering.
Kondaveeti, Hari Kishan, Kumaravelu, Nandeesh Kumar, Vanambathina, Sunny Dayal, Mathe, Sudha Ellison, and Vappangi, Suseela
Computer Science Review. May, 2021, Vol. 40
Keywords Raspberry Pi; BeagleBone; Sharks Cove; Waspmote Abstract Arduino, an open-source electronics platform, has become the go-to option for anyone working on interactive hardware and software projects. An Arduino board (such as the Uno) connected to a breadboard with plugins such as inputs, sensors, lights, and displays can be controlled by a code written in the Arduino development environment. How to achieve this is by prototyping with Arduino. Prototyping with Arduino has grown in popularity with the increased use of the Arduino platform. Prototyping with Arduino, however, is not an easy task for nonprogrammers with interest in the field. With increased public interest in the field will come a need for accessible information. This paper presents a methodical literature review intended to intensively analyze and compare existing primary studies on prototyping with Arduino. We found about 130 of such studies, all peer-reviewed and published within the last 15 years, including these years (2015--2020). These studies were tediously and carefully chosen through a three-step process. In this paper, a cautious analysis of selected studies was followed by a clear description of the methods applied. The methods were categorized according to the success rate of the studied prototypes. Results obtained can be used in researches on the best technique to adopt while prototyping with Arduino. They can also be used in electronics researches and by individuals who wish to obtain a guide on prototyping with Arduino despite lacking grounded knowledge of the subject matter. Author Affiliation: (a) School of Computer Science & Engineering, VIT-AP University, Beside AP Secretariat, Near Vijayawada, Andhra Pradesh, India (b) School of Electronics Engineering, VIT-AP University, Beside AP Secretariat, Near Vijayawada, Andhra Pradesh, India * Corresponding author. Article History: Received 22 September 2020; Accepted 13 January 2021 Byline: Hari Kishan Kondaveeti [firstname.lastname@example.org] (a,*), Nandeesh Kumar Kumaravelu (b), Sunny Dayal Vanambathina (b), Sudha Ellison Mathe (b), Suseela Vappangi (b)
Hosic S, Bindas AJ, Puzan ML, Lake W, Soucy JR, Zhou F, Koppes RA, Breault DT, Murthy SK, and Koppes AN
ACS biomaterials science & engineering [ACS Biomater Sci Eng] 2021 Jul 12; Vol. 7 (7), pp. 2949-2963. Date of Electronic Publication: 2020 Jun 03.
Microfluidic organs-on-chips aim to realize more biorelevant in vitro experiments compared to traditional two-dimensional (2D) static cell culture. Often such devices are fabricated via poly(dimethylsiloxane) (PDMS) soft lithography, which offers benefits (e.g., high feature resolution) along with drawbacks (e.g., prototyping time/costs). Here, we report benchtop fabrication of multilayer, PDMS-free, thermoplastic organs-on-chips via laser cut and assembly with double-sided adhesives that overcome some limitations of traditional PDMS lithography. Cut and assembled chips are economical to prototype ($2 per chip), can be fabricated in parallel within hours, and are Luer compatible. Biocompatibility was demonstrated with epithelial line Caco-2 cells and primary human small intestinal organoids. Comparable to control static Transwell cultures, Caco-2 and organoids cultured on chips formed confluent monolayers expressing tight junctions with low permeability. Caco-2 cells-on-chip differentiated ∼4 times faster, including increased mucus, compared to controls. To demonstrate the robustness of cut and assemble, we fabricated a dual membrane, trilayer chip integrating 2D and 3D compartments with accessible apical and basolateral flow chambers. As proof of concept, we cocultured a human, differentiated monolayer and intact 3D organoids within multilayered contacting compartments. The epithelium exhibited 3D tissue structure and organoids expanded close to the adjacent monolayer, retaining proliferative stem cells over 10 days. Taken together, cut and assemble offers the capability to rapidly and economically manufacture microfluidic devices, thereby presenting a compelling fabrication technique for developing organs-on-chips of various geometries to study multicellular tissues.
The growing need for the implementation of stretchable biosensors in the body has driven rapid prototyping schemes through the direct ink writing of multidimensional functional architectures. Recent approaches employ biocompatible inks that are dispensable through an automated nozzle injection system. However, their application in medical practices remains challenged in reliable recording due to their viscoelastic nature that yields mechanical and electrical hysteresis under periodic large strains. Herein, we report sponge-like poroelastic silicone composites adaptable for high-precision direct writing of custom-designed stretchable biosensors, which are soft and insensitive to strains. Their unique structural properties yield a robust coupling to living tissues, enabling high-fidelity recording of spatiotemporal electrophysiological activity and real-time ultrasound imaging for visual feedback. In vivo evaluations of custom-fit biosensors in a murine acute myocardial infarction model demonstrate a potential clinical utility in the simultaneous intraoperative recording and imaging on the epicardium, which may guide definitive surgical treatments.