Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR or more precisely CRISPR-Cas) system has proven to be a highly efficient and simple tool for achieving site-specific genome modifications in comparison to Zinc Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs). The discovery of bacterial defense system that uses RNA-guided DNA cleaving enzymes for producing double-strand breaks along CRISPR has provided an exciting alternative to ZFNs and TALENs for gene editing & regulation, as the CRISPR-associated (Cas) proteins remain the same for different gene targets and only the short sequence of the guide RNA needs to be changed to redirect the site-specific cleavage. Therefore, in recent years the CRISPR-Cas system has emerged as a revolutionary engineering tool for carrying out precise and controlled genetic modifications in many microbes such as Escherichia coli, Staphylococcus aureus, Lactobacillus reuteri, Clostridium beijerinckii, Streptococcus pneumonia, and Saccharomyces cerevisiae. Though, concerns about CRISPR-Cas effectiveness in interlinked gene modifications and off-target effects need to be addressed. Nevertheless, it holds a great potential to speed up the pace of gene function discovery by interacting with previously intractable organisms and by raising the extent of genetic screens. Therefore, the potential applications of this system in microbial adaptive immune system, genome editing, gene regulations, functional genomics & biosynthesis along ethical issues, and possible harmful effects have been reviewed.
Near-infrared fluorescent dyes based on small organic molecules are believed to have a great influence on cancer diagnosis at large and targeted cancer cell bioimaging, in particular. NIR dyes-based organic molecules have notable characteristics features, such as high tissue penetration and low tissue autofluorescence in the NIR spectral region. Cancer targeted bioimaging relies significantly on the synthesis of highly specific molecular probes with excellent stability. Recently, NIR dyes have emerged as unique fluorescent probes for cancer bioimaging. These current advancements have overcome many limitations of conventional NIR probes e.g., poor photostability and hydrophilicity, insufficient stability and low quantum yield. The further potential lies in NIR dyes or NIR dyes-coated nanocarriers conjugated with cancer-specific ligand (e.g., peptides, antibodies, proteins or other small molecules). Multifunctional NIR dyes have synthesized, which efficiently accumulate in cancer cells without requiring chemical conjugation and also these dyes have presented novel photophysical and pharmaceutical properties for in vivo imaging. This review highlights the recently developed NIR dyes with novel applications in cancer bioimaging. We believe that these novel fluorophores will enhance our understanding of cancer imaging and pave a new road in cancer diagnosis and treatment.
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