ABSTRACT
Precise genetic engineering in specific cell types within an intact organism is intriguing yet challenging, especially in a spatiotemporal manner without the interference caused by chemical inducers. Here we engineered a photoactivatable Dre recombinase based on the identification of an optimal split site and demonstrated that it efficiently regulated transgene expression in mouse tissues spatiotemporally upon blue light illumination. Moreover, through a double-floxed inverted open reading frame strategy, we developed a Cre-activated light-inducible Dre (CALID) system. Taking advantage of well-defined cell-type-specific promoters or a well-established Cre transgenic mouse strain, we demonstrated that the CALID system was able to activate endogenous reporter expression for either bulk or sparse labeling of CaMKIIα-positive excitatory neurons and parvalbumin interneurons in the brain. This flexible and tunable system could be a powerful tool for the dissection and modulation of developmental and genetic complexity in a wide range of biological systems.
Subject(s)
Escherichia coli Proteins/metabolism , Genetic Engineering , Genome , Light , Recombinases/metabolism , Animals , Brain/metabolism , Dependovirus/metabolism , Gene Expression , Genes, Reporter , Genetic Vectors/metabolism , HEK293 Cells , Humans , Integrases/metabolism , Liver/metabolism , Mice, Inbred C57BL , Mice, Transgenic , Neurons/metabolism , Time FactorsABSTRACT
BACKGROUND: Shrimp is widely consumed around the world. Since muscle is the primary edible component of shrimp, muscle quality (particularly texture) has a direct impact on the economic value of shrimp products. However, reports on the shrimp muscle quality influenced by transportation are rather limited, and the underlying mechanism remains unknown. RESULTS: During the simulated transportation, the water pH and total ammonia-nitrogen content and un-ionized ammonia contents were elevated. Furthermore, reductions in shrimp muscle water-holding capacity, hardness, and shear value with intensive myofibrillar protein degradation were detected. Simulated transportation decreased the pH and glycogen content of shrimp muscle while increasing lactic dehydrogenase activity and lactate content, resulting in an elevated level of free calcium ions and increased µ-calpain and general proteolytic activities. Water exchange could improve the water quality and reduce the mortality of shrimp during transportation, as well as decrease muscle textural softening by alleviating these stress responses. CONCLUSIONS: Maintaining water quality and, in particular, reducing ammonia are critical to improving shrimp survival and muscle quality during live transportation. This study is of great significance for the better maintenance of the textural properties of shrimp meat. © 2023 Society of Chemical Industry.
Subject(s)
Ammonia , Penaeidae , Animals , Penaeidae/chemistry , Seafood , Nitrogen , MusclesABSTRACT
Protein-starch interaction has an important impact on the properties of starchy foods rich in protein, but the contribution of the interaction to Chinese yam still remains unclear. This study aimed to characterize the physicochemical and functional properties related to the possible interaction between starch and protein in Chinese yam. Differential scanning calorimetry and rapid viscosity analyzer results revealed that the gelatinization temperature increased in protein and starch cross-linked powder, while the peak viscosity and the setback viscosity decreased. The swelling power and solubility at 80°C and 95°C decreased with increasing protein ratio in the powder. In vitro starch digestibility test indicated that a high protein ratio could rapidly reduce digestible starch, but increase both slowly digestible starch and resistant starch. Protein could act as the physical barrier toward starch against heating and digestion to exert the influence on starch properties. Fourier transform infrared spectroscopy test revealed the interaction between protein and starch. These results revealed the role of protein-starch interaction and provided beneficial information for the utilization of Chinese yam.
ABSTRACT
Optogenetic genome engineering is a powerful technology for high-resolution spatiotemporal genetic manipulation, especially for in vivo studies. It is difficult to generate stable transgenic animals carrying a tightly regulated optogenetic system, as its long-term expression induces high background activity. Here, the generation of an enhanced photoactivatable Cre recombinase (ePA-Cre) transgenic mouse strain with stringent light responsiveness and high recombination efficiency is reported. Through serial optimization, ePA-Cre is developed to generate a transgenic mouse line that exhibits 175-fold induction upon illumination. Efficient light-dependent recombination is detected in embryos and various adult tissues of ePA-Cre mice crossed with the Ai14 tdTomato reporter. Importantly, no significant background Cre activity is detected in the tested tissues except the skin. Moreover, efficient light-inducible cell ablation is achieved in ePA-Cre mice crossed with Rosa26-LSL-DTA mice. In conclusion, ePA-Cre mice offer a tightly inducible, highly efficient, and spatiotemporal-specific genome engineering tool for multiple applications.