CRISPR-based assays for rapid detection of SARS-CoV-2.

COVID-19 pandemic posed an unprecedented threat to global public health and economies. There is no effective treatment of the disease, hence, scaling up testing for rapid diagnosis of SARS-CoV-2 infected patients and quarantine them from healthy individuals is one the best strategies to curb the pandemic. Establishing globally accepted easy-to-access diagnostic tests is extremely important to understanding the epidemiology of the present pandemic. While nucleic acid based tests are considered to be more sensitive with respect to serological tests but present gold standard qRT-PCR-based assays possess limitations such as low sample throughput, requirement for sophisticated reagents and instrumentation. To overcome these shortcomings, recent efforts of incorporating LAMP-based isothermal detection, and minimizing the number of reagents required are on rise. CRISPR based novel techniques, when merge with isothermal and allied technologies, promises to provide sensitive and rapid detection of SARS-CoV-2 nucleic acids. Here, we discuss and present compilation of state-of-the-art detection techniques for COVID-19 using CRISPR technology which has tremendous potential to transform diagnostics and epidemiology.

Year-round sustainable biomass production potential of Nannochloris sp. in outdoor raceway pond enabled through strategic photobiological screening.

Microalgae cultivation utilizes the energy of sunlight to reduce carbon dioxide (CO2) for producing renewable energy feedstock. The commercial success of the biological fixation of carbon in a consistent manner depends upon the availability of a robust microalgae strain. In the present work, we report the identification of a novel marine Nannochloris sp. through multiparametric photosynthetic evaluation. Detailed photobiological analysis of this strain has revealed a smaller functional antenna, faster relaxation kinetics of non-photochemical quenching, and a high photosynthetic rate with increasing light and temperatures. Furthermore, laboratory scale growth assessment demonstrated a broad range halotolerance of 10-70 parts per thousand (PPT) and high-temperature tolerance up to 45 °C. Such traits led to the translation of biomass productivity potential from the laboratory scale (0.2-3.0 L) to the outdoor 50,000 L raceway pond scale (500-m2) without any pond crashes. The current investigation revealed outdoor single-day peak areal biomass productivity of 43 g m-2 d-1 in summer with an annual (March 2019-February 2020) average productivity of 20 g m-2 d-1 in seawater. From a sustainability perspective, this is the first report of successful round-the-year (> 347 days) multi-season (summer, monsoon, and winter) outdoor cultivation of Nannochloris sp. in broad seawater salinity (1-57 PPT), wide temperature ranges (15-40 °C), and in fluctuating light conditions. Concurrently, outdoor cultivation of this strain demonstrated conducive fatty acid distribution, including increased unsaturated fatty acids in winter. This inherent characteristic might play a role in protecting photosynthesis machinery at low temperatures and in high light stress. Altogether, our marine Nannochloris sp. showed tremendous potential for commercial scale cultivation to produce biofuels, food ingredients, and a sustainable source for vegetarian protein.

Whole genome sequence analysis of Geitlerinema sp. FC II unveils competitive edge of the strain in marine cultivation system for biofuel production.

A filamentous cyanobacteria, Geitlerinema sp. FC II, was isolated from marine algae culture pond at Reliance Industries Limited (RIL), India. The 6.7 Mb draft genome of FC II encodes for 6697 protein coding genes. Analysis of the whole genome sequence revealed presence of nif gene cluster, supporting its capability to fix atmospheric nitrogen. FC II genome contains two variants of sulfide:quinone oxidoreductases (SQR), which is a crucial elector donor in cyanobacterial metabolic processes. FC II is characterized by the presence of multiple CRISPR- Cas (Clustered Regularly Interspaced Short Palindrome Repeats - CRISPR associated proteins) clusters, multiple variants of genes encoding photosystem reaction centres, biosynthetic gene clusters of alkane, polyketides and non-ribosomal peptides. Presence of these pathways will help FC II in gaining an ecological advantage over other strains for biomass production in large scale cultivation system. Hence, FC II may be used for production of biofuel and other industrially important metabolites.

Enhanced Algal Photosynthetic Photon Efficiency by Pulsed Light.

We present experimental results demonstrating that, relative to continuous illumination, an increase of a factor of 3-10 in the photon efficiency of algal photosynthesis is attainable via the judicious application of pulsed light for light intensities of practical interest (e.g., average-to-peak solar irradiance). We also propose a simple model that can account for all the measurements. The model (1) reflects the essential rate-limiting elements in bioproductivity, (2) incorporates the impact of photon arrival-time statistics, and (3) accounts for how the enhancement in photon efficiency depends on the timescales of light pulsing and photon flux density. The key is avoiding "clogging" of the photosynthetic pathway by properly timing the light-dark cycles experienced by algal cells. We show how this can be realized with pulsed light sources, or by producing pulsed-light effects from continuous illumination via turbulent mixing in dense algal cultures in thin photobioreactors.

CRISPR-Cas9 System for Genome Engineering of Photosynthetic Microalgae.

Targeted genome editing using RNA-guided endonucleases is an emerging tool in algal biotechnology. Recently, CRISPR-Cas systems have been widely used to manipulate the genome of some freshwater and marine microalgae. Among two different classes, and six distinct types of CRISPR systems, Cas9-driven type II system has been widely used in most of the studies for targeted knock-in, knock-out and knock-down of desired genes in algae. CRISPR technology has been demonstrated in microalgae including diatoms to manifest the function of the particular gene (s) and developing industrial traits, such as improving lipid content and biomass productivity. Instead of these, there are a lot of gears to be defined about improving efficiency and specificity of targeted genome engineering of microalgae using CRISPR-Cas system. Optimization of tools and methods of CRISPR technology can undoubtedly transform the research toward the industrial-scale production of commodity chemicals, food and biofuels using photosynthetic cell factories. This review has been focused on the efforts made so far to targeted genome engineering of microalgae, identified scopes about the hurdles related to construction and delivery of CRISPR-Cas components, algae transformation toolbox, and outlined the future prospect toward developing the CRISPR platform for high-throughput genome-editing of microalgae.

Algae to Economically Viable Low-Carbon-Footprint Oil.

Algal oil as an alternative to fossil fuel has attracted attention since the 1940s, when it was discovered that many microalgae species can produce large amounts of lipids. Economics and energy security were the motivational factors for a spurt in algae research during the 1970s, 1990s, and early 2000s. Whenever crude prices declined, research on algae stopped. The scenario today is different. Even given low and volatile crude prices ($30-$50/barrel), interest in algae continues all over the world. Algae, with their cure-all characteristics, have the potential to provide sustainable solutions to problems in the energy-food-climate nexus. However, after years of effort, there are no signs of algae-to-biofuel technology being commercialized. This article critically reviews past work; summarizes the current status of the technology; and based on the lessons learned, provides a balanced perspective on a potential path toward commercialization of algae-to-oil technology.