Microbes use diverse defence strategies that allow them to withstand exposure to a variety of genome invaders such as bacteriophages and plasmids. One such defence strategy is the use of RNA guided endonuclease called CRISPR-associated (Cas) 9 protein. The Cas9 protein, derived from type II CRISPR/Cas system, has been adapted as a versatile tool for genome targeting and engineering due to its simplicity and high efficiency over the earlier tools such as ZFNs and TALENs. With recent advancements, CRISPR/Cas9 technology has emerged as a revolutionary tool for modulating the genome in living cells and inspires innovative translational applications in different fields. In this paper we review the developments and its potential uses in the CRISPR/Cas9 technology as well as recent advancements in genome engineering using CRISPR/Cas9.

Plants have long been considered as a basis of medicines for different indigenous cultures around the globe. They continue as a prominent source of important phytoconstituents which exhibit significant biological activities. In this study, we performed the phytochemical screening, estimation of total phenolic and flavonoids, antioxidants, and antimicrobial activities of the stem bark of Beilschmiedia roxburghiana Nees using different solvents. The total phenolic and total flavonoid contents ranged from 106.73±1.62mg GAE/g and 99.32±0.66mg QE/g (methanol extract) to 65.59±1.79mg GAE/g and 29.98±0.90mg QE/g (n-hexane extract), respectively. The maximum 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity with a half-maximal inhibitory concentration (IC₅₀) of 39.86±3.69μg/mL was observed for methanol extract followed by aqueous (IC₅₀=43.55±6.16μg/mL), ethyl acetate (IC₅₀=44.30±5.88μg/mL), dichloromethane (IC₅₀=71.50±4.70μg/mL), and the lowest activity was observed for n-hexane extract. The disc diffusion method revealed that the ethyl acetate extract exhibited relatively higher activity against Salmonella typhi (ZOI=13mm), and moderate activities against Shigella sonnei, Acinetobacter baumannii, Klebsiella pneumoniae, and Staphylococcus aureus (ZOI=12mm). The methanol and aqueous extracts showed nearly parallel and the n-hexane and dichloromethane extracts exhibited mild antibacterial activities. The results indicated that the polarity index of the extracting solvents amplified the biological activities of the extract. The study is helpful to support the validity of the traditional application of the plant as natural medicine.

Breast cancer is one of the most common cancers with a high mortality rate, underscoring the need to identify new therapeutic targets. Here we report that non-POU domain-containing octamer-binding (NONO) protein is overexpressed in breast cancer and validated the interaction of the WW domain of PIN1 with c-terminal threonine-proline (thr-pro) motifs of NONO. The interaction of NONO with PIN1 increases the stability of NONO by inhibiting its proteasomal degradation, and this identifies PIN1 as a positive regulator of NONO in promoting breast tumor development. Functionally, silencing of NONO inhibits the growth, survival, migration, invasion, epithelial to mesenchymal transition (EMT), and stemness of breast cancer cells in vitro. A human metastatic breast cancer cell xenograft was established in transparent zebrafish (Danio rerio) embryos to study the metastatic inability of NONO-silenced breast cancer cells in vivo. Mechanistically, NONO depletion promotes the expression of the PDL1 cell-surface protein in breast cancer cells. The identification of novel interactions of NONO with c-Jun and β-catenin proteins and activation of the Akt/MAPK/β-catenin signaling suggests that NONO is a novel regulator of Akt/MAPK/β-catenin signaling pathways. Taken together, our results indicated an essential role of NONO in the tumorigenicity of breast cancer and could be a potential target for anti-cancerous drugs.

This review traces the evolution of clustered regularly interspaced short palindromic repeats (CRISPR) technology from a prokaryotic immune mechanism to a versatile tool for precise genome engineering. We compare CRISPR with traditional gene-editing methods like RNA interference (RNAi), zinc finger nucleases (ZFNs), and transcription activator-like effector nucleases (TALENs), emphasizing its advantages in target specificity, multiplexing, and ease of design. We examine various Cas enzyme classes, engineered variants, and their applications in dissecting genetic alterations at the cellular level. The review further explores CRISPR’s expanding role in developing disease models using tissues, organoids, and animal systems, enhancing our understanding of disease mechanisms. Finally, we discuss CRISPR’s emerging applications in diagnostics and its transformative impact on immunotherapy and cell-based cancer treatments.