CTX-1(MiRx Pharmaceuticals)

Osteoporosis is the deterioration of bone mineral density (BMD) because of an imbalance between bone resorption and formation, which might happen due to lots of factors like age, hormonal imbalance, and several others. While this occurrence is prevalent in both genders, it is more common in women, especially postmenopausal women. It is an asymptomatic disease that is underlying until the first incidence of a fracture. The bone is weakened, making it more susceptible to fracture. Even a low trauma can result in a fracture, making osteoporosis an even more alarming disease. These fractures can sometimes be fatal or can make the patient bedridden. Osteoporosis is an understudied disease, and there are certain limitations in diagnosing and early-stage detection of this condition. The standard method of dual X-ray absorptiometry can be used to some extent and can be detected in standard radiographs after the deterioration of a significant amount of bone mass. Clinically assessing osteoporosis using biomarkers can still be challenging, as clinical tests can be expensive and cannot be accessed by most of the general population. In addition, manufacturing antibodies specific to these biomarkers can be a challenging, time-consuming, and expensive method. As an alternative to these antibodies, molecularly imprinted polymers (MIPs) can be used in the detection of these biomarkers. This Review provides a comprehensive exploration of bone formation, resorption, and remodeling processes, linking them to the pathophysiology of osteoporosis. It details biomarker-based detection and diagnosis methods, with a focus on MIPs for sensing CTX-1, NTX-1, and other biomarkers. The discussion compares traditional clinical practices with MIP-based sensors, revealing comparable sensitivity with identified limitations. Additionally, the Review contrasts antibody-functionalized sensors with MIPs. Finally, our Review concludes by highlighting the potential of MIPs in future early-stage osteoporosis detection.

CRISPR-Cas9-mediated genome editing in sheep is of great use in both agricultural and biomedical applications. While targeted gene knockout by CRISPR-Cas9 through non-homologous end joining (NHEJ) has worked efficiently, the knockin efficiency via homology-directed repair (HDR) remains lower, which severely hampers the application of precise genome editing in sheep. Here, in sheep fetal fibroblasts (SFFs), we optimized several key parameters that affect HDR, including homology arm (HA) length and the amount of double-stranded DNA (dsDNA) repair template; we also observed synchronization of SFFs in G2/M phase could increase HDR efficiency. Besides, we identified three potent small molecules, RITA, Nutlin3, and CTX1, inhibitors of p53-MDM2 interaction, that caused activation of the p53 pathway, resulting in distinct G2/M cell-cycle arrest in response to DNA damage and improved CRISPR-Cas9-mediated HDR efficiency by 1.43- to 4.28-fold in SFFs. Furthermore, we demonstrated that genetic knockout of p53 could inhibit HDR in SFFs by suppressing the expression of several key factors involved in the HDR pathway, such as BRCA1 and RAD51. Overall, this study offers an optimized strategy for the usage of dsDNA repair template, more importantly, the application of MDM2 antagonists provides a simple and efficient strategy to promote CRISPR/Cas9-mediated precise genome editing in sheep primary cells.