The genetically modified human foreskin fibroblast cell line (YhFF#8) stably expressing Cas9 gene: A lab resource report.

Background: Stable Cas9 (CRISPR-associated protein 9)-expressing cell lines have emerged as valuable tools in genetic research, enhancing the efficiency of the CRISPR/Cas9 system and streamlining gene editing procedures. These cell lines enable simultaneous editing of multiple genes and reduce the overall editing time. Objective: This study aimed to develop a stable human fibroblast cell line capable of genetic conversion into a mutant form, serving as a cellular model for a specific genetic disease. The established cell line facilitates investigation of disease mechanisms, testing of potential treatments, and gaining insights into underlying molecular processes. Materials and Methods: Human embryonic kidney 293LTV cells were used to produce pseudo-virus particles, while Yazd human foreskin fibroblasts batch 8 (YhFF#8) cells were targeted for genetic modification. Transfection of human embryonic kidney 293LTV cells with pCDH-Cas9 plasmid DNA generated pseudo-viral particles. YhFF#8 cells were transduced and selected using antibiotics. Green fluorescent protein (GFP) detection confirmed successful transduction and selection. Relative expression levels of the Cas9 gene were determined by quantitative polymerase chain reaction. Results: The study validated the fidelity of the Cas9 gene cassette sequence and its transcriptional activity. Transduced YhFF#8 cells exhibited green fluorescence, with antibiotic selection resulting in nearly 100% transduced cells. A reporter GFP gene enabled real-time monitoring of YhFF#8-Cas9-GFP-PuroR cells using fluorescence microscopy. Conclusion: YhFF#8-Cas9-GFP-PuroR cells, labeled and susceptible to genomic editing, provide an optimal source for generating induced pluripotent stem cell lines for future biomedical research.

Does combination of estradiol and sesame oil improve the oocyte quality, embryo development and expressions of Zp3, E-cad, and Ctnnb1 genes in mice? An experimental study.

BACKGROUND: Aging may reduce oocyte maturation, embryo quality, and fertility potential. OBJECTIVE: To compare the effect of estradiol (E2) and sesame oil on oocyte and embryo quality between young and old mice. MATERIALS AND METHODS: Sixty old and young female mice were divided in to two groups (30 mice/group, grouped by age). Each group was divided into three subgroups of mice treated with sesame oil, E2 + sesame oil, and normal saline as control group. After ovulation induction, some oocytes were considered for in vitro fertilization and the rest were assessed for morphological status. After obtaining the two-cell embryos, the embryos were collected to determine the expression of zona pellucida (ZP) glycoprotein 3, E-cadherin, and ß-catenin genes and some of them followed until the blastocysts stage to evaluate the viability. RESULTS: The findings showed that the mean ZP and perivitelline space thickness increased in the old mice that received the E2 + sesame oil treatment. The number of 2-cell embryos, blastocysts, and live cells were significantly higher in the old group treated with sesame oil respectively (p = 0.018, 0.002, and < 0.0001, respectively). The normal ZP shape and refractile body numbers increased in the old mice that were treated with sesame oil, respectively. The E-cadherin gene was downregulated in the treatment groups compared to the controls. CONCLUSION: Sesame oil showed a better response in the old mice, because aging is associated with an increased rate of reactive oxygen species, causing deficiencies in both oocyte and embryo qualities.

Association of rs670 variant of APOA-1 gene with cardiometabolic markers after consuming sesame, canola and sesame-canola oils in adults with and without type 2 diabetes mellitus.

BACKGROUND & AIMS: The inter-individual variations of the metabolic markers in response to dietary interventions may be mediated by genetic factors. We examined whether the type of dietary oils can modulate the effects of -75G/A polymorphism in APOA-1 gene on cardiometabolic markers. METHODS: This study was a randomized, triple-blind, cross-over clinical trial. Participants with and without type 2 diabetes were randomly assigned to replace their regular oil with sesame oil, canola oil and sesame-canola oil for 9 weeks. Genotyping was conducted using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: Ninety-five diabetes patients and 73 healthy individuals completed the study protocol. In patients with type 2 diabetes, the A allele carriers experienced greater decrease in systolic blood pressure compared with GG homozygotes following sesame-canola oil intake. Serum levels of HDL-C and TG: HDL ratio was increased and decreased following canola oil intake in patients carrying the A allele rather than non-A allele carriers, respectively. More reductions for risk of cardiovascular diseases and mortality, except risk of stroke were found in the A allele carriers compared with GG homozygotes after intakes of canola and sesame-canola oils, but not sesame oil. There was also a significant genotype effect as well as genotype-dietary oil interactions on cardiovascular risk scores. In healthy individuals, a considerable decrease in visceral fat was accompanied by a significant increase in HDL-C levels in the A allele carriers compared with non-A allele carriers after sesame oil intake. CONCLUSION: Patients with diabetes carrying the A allele might benefit from canola and sesame-canola oils intakes, and healthy A allele carriers from sesame and sesame-canola oils intakes as well. Future clinical trials are recommended to warrant current findings.

The combined effects of cholesteryl ester transfer protein (CETP) TaqIB gene polymorphism and canola, sesame and sesame-canola oils consumption on metabolic response in patients with diabetes and healthy people.

Introduction: Cholesteryl ester transfer protein (CETP) is a key regulating enzyme in the lipid metabolism pathway, and its gene polymorphism may be a candidate for modulating the metabolic responses to dietary intervention. We thus examined whether the effects of the CETP TaqIB polymorphism on metabolic profiles were modified by dietary plant oils. Methods: This is a retrospective analysis of data collected during a randomized triple-blind cross over trial. A total of 95 patients with type 2 diabetes and 73 non-diabetes individuals completed a 9-weekof the intake of sesame, canola and sesame-canola oils. Blood samples were collected at the beginning and at the end of each intervention period for biochemical analysis. Genotyping was done using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: In diabetes patients, B1B1 homozygotes of the CETP TaqIB polymorphism compared with B2 carriers (B1B2 + B2B2) had significantly lower diastolic blood pressure, apoB and apoB: apoA-1,and higher Lp(a) after the intake of sesame-canola oil, as well as lower insulin and HOMA-IR after the intake of sesame oil. There was also a significant effect of genotype on adjusted changes of apoB, apoB: apoA-1, insulin, HOMA-IR and QUICKI. A significant genotype-dietary oils combined effects were observed for diastolic blood pressure, and LDL: HDL, TC: HDL and TG: HDL ratios in diabetes patients. No independent or combined effects of dietary oils and genotypes on outcomes were found in healthy people. Conclusion: There was a modulatory effect of the CETP TaqIB polymorphism on some metabolic traits in response to plant oils in patients with diabetes. Taken together, the intake of sesame-canola and canola oils showed more favorable effects in diabetes patients with B1B1 genotype. Future investigations are needed to confirm these results.

Nano-carbohydrates: Synthesis and application in genetics, biotechnology, and medicine.

Combining nanoparticles with carbohydrate has triggered an exponential growth of research activities for the design of novel functional bionanomaterials, nano-carbohydrates. Recent advances in versatile synthesis of glycosylated nanoparticles have paved the way towards diverse biomedical applications. The accessibility of a wide variety of these structured nanosystems, in terms of shape, size, and organization around stable nanoparticles, has readily contributed to their development and application in nanomedicine. Glycosylated gold nanoparticles, glycosylated quantum dots, fullerenes, single-wall nanotubes, and self-assembled glyconanoparticles using amphiphilic glycopolymers or glycodendrimers have received considerable attention for their application in powerful imaging, therapeutic, and biodiagnostic devices. Recently, nano-carbohydrates were used for different types of microarrays to detect proteins and nucleic acids.