CRISPR/Cas9-Mediated Gene Editing in Salmonids Cells and Efficient Establishment of Edited Clonal Cell Lines.

Finfish production has seen over three-fold increase in the past 30 years (1990-2020), and Atlantic salmon (A. salmon; salmo salar) accounted for approximately 32.6% of the total marine and coastal aquaculture of all finfish species in the year 2020, making it one of the most profitable farmed fish species globally. This growth in production is, however, threatened by a number of problems which can be solved using the CRISPR/Cas technology. In vitro applications of CRISPR/Cas using cell lines can complement its in vivo applications, but salmonids-derived cell lines are difficult to gene edit because they grow slowly, are difficult to transfect and isolate single clones of gene-edited cells. While clonal isolation of the gene-edited Chinook salmon cell line (CHSE-214) has successfully been performed, there is no report of successful clonal isolation of the gene-edited A. salmon ASK-1 and SHK-1cell lines. In the current study, two gene loci-cr2 and mmp9 of A. salmon-were efficiently edited using the ribonucleoprotein (RNP) and plasmid CRISPR/Cas9 strategies. Edited cells were enriched using flow cytometer-activated cell sorting (FACS), followed by clonal isolation and expansion of edited cells. The study both confirms the recent report of the highly efficient editing of these widely used model cell lines, as well as extends the frontline in the single-cell cloning of gene-edited salmonids cells. The report also highlights the pitfalls and future directions in the application of CRISPR/Cas9 in these cells.

Risk of Atrial Fibrillation in Pemphigus Vulgaris.

Background: Pemphigus vulgaris (PV) is a rare immunobullous disease with a higher mortality rate than the general population. The aim of this study was to investigate P-wave duration and P-wave dispersion (PWD) in patients with PV as predictors of atrial fibrillation (AF). Materials and Methods: In this case-control study, the risk of AF development was determined by measuring maximum and minimum P-wave duration (P-max and P-min) and PWD in 45 PV patients and 45 healthy individuals. The incidence of metabolic syndrome was evaluated. Results: PWD and P-max values of the study group were significantly higher than those of the control group. No difference was observed between PWD with regards to disease duration and disease phenotype (p > 0.05). There was no significant difference regarding the prevalence of metabolic syndrome in PV patients compared with the control group. Conclusion: PWD and P-max, which are accepted as risk factors for AF development, were found to be higher in PV patients. Some components of metabolic syndrome were more prevalent in PV patients. It seems that there is an increased risk of CVD and AF in PV patients.