Biochemical assays articles from across Nature Portfolio

Fusion proteins between roGFP2 and glutaredoxins are used for intracellular redox measurements. Here, the authors determined all rate constants of the reaction cycle for roGFP2 measurements and identified an alternative glutaredoxin mechanism.

The coenzyme Q biosynthetic pathway has evaded full characterization for decades, in part due to the inherent insolubility of coenzyme Q and the instability of its membrane-associated biosynthetic enzymes. Now, researchers have resurrected an active ancestral coenzyme Q metabolon in vitro that has unveiled valuable insights into previously uncharacterized aspects of coenzyme Q biosynthesis.

Combining the precision of CRISPR’s DNA searching ability with the speed and scalability of electronics, we have developed an ‘electronic DNA search engine’, called a CRISPR–Chip, which not only enables DNA detection without amplification, but also showcases the untapped potential of merging molecular biology with nanomaterial electronics. Here, we discuss highlights and challenges on the journey from the initial idea to the commercialization of the CRISPR–Chip.

We developed exponentially amplified rolling circle amplification with CRISPR–Cas12a as a one-pot, isothermal assay for microRNA detection. This method has single-digit femtomolar sensitivity and single-nucleotide specificity, and can be deployed for point-of-care testing. The assay has been adapted for the microRNA profiling of extracellular vesicles, which is used in the diagnosis of pancreatic cancer.

CRISPR-based assays can be adopted as ultrasensitive molecular diagnostics in resource-limited settings, but point-of-care applications must address additional requirements. Here, we discuss the major obstacles for developing these assays and offer insights into how to surmount them.

Plant cell wall pectin has a homogalacturonan or rhamnogalacturonan backbone. Incomplete knowledge of RG-I biosynthetic enzymes has impeded in vitro pectin synthesis and pectin structure and function studies. Here, RGGAT1 is identified as a GT116 RG-I backbone biosynthetic galacturonosyltransferase that produces polymeric RG-I backbone when expressed with GT106 RG-I rhamnosyltransferases.

Organic electrochemical transistors functionalized with antigen-specific nanobodies can rapidly and specifically detect antigens at attomolar-to-nanomolar levels in bodily fluids.