CRISPR/Cas9 systems:Delivery technologies and biomedical applications

The emergence of the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)genome-editing system has brought about a significant revolution in the realm of managing human diseases,establishing animal models,and so on.To fully harness the potential of this potent gene-editing tool,ensuring efficient and secure delivery to the target site is paramount.Consequently,developing effective delivery methods for the CRISPR/Cas9 system has become a critical area of research.In this review,we present a comprehensive outline of delivery strategies and discuss their biomedical applications in the CRISPR/Cas9 system.We also provide an indepth analysis of physical,viral vector,and non-viral vector delivery strategies,including plasmid-,mRNA-and protein-based approach.In addition,we illustrate the biomedical applications of the CRISPR/Cas9 system.This review highlights the key factors affecting the delivery process and the current challenges facing the CRISPR/Cas9 system,while also delineating future directions and prospects that could inspire innovative delivery strategies.This review aims to provide new insights and ideas for advancing CRISPR/Cas9-based delivery strategies and to facilitate breakthroughs in biomedical research and therapeutic applications.

The role of circadian clocks in cancer:Mechanisms and clinical implications

Circadian rhythm refers to the inherent 24-h cycle oscillation of biochemical,phys-iological and behavioral functions,which is almost universal in eukaryotes.At least 14 core clock genes have been reported to form multiple chain feedback loops that confer intrinsic circadian rhythmicity onto the molecular clock.Accumulating evidence has shown that the circadian gene dysfunction resulted from single nucleotide polymorphisms(SNPs),deletions,epigenetic modification,and deregulation is strongly associated with cancer risk.In the pre-sent review,we describe the composition of circadian rhythm system.We highlight the func-tion and mechanism of clock genes in cancer pathogenesis and progression.Moreover,their potential clinical implications as prognostic biomarkers and therapeutic targets have been ad-dressed.

Drp1-dependent remodeling of mitochondrial morphology triggered by EBV-LMP1 increases cisplatin resistance

Latent membrane protein 1(LMP1)is a major Epstein–Barr virus(EBV)-encoded oncoprotein involved in latency infection that regulates mitochondrial functions to facilitate cell survival.Recently,mitochondrial fission has been demonstrated as a crucial mechanism in oncovirus-mediated carcinogenesis.Mitochondrial dynamin-related protein 1(Drp1)-mediated mitochondrial fission has an impact on the chemoresistance of cancers.However,the mechanism by which oncogenic stress promotes mitochondrial fission,potentially contributing to tumorigenesis,is not entirely understood.The role of Drp1 in the oncogenesis and prognosis of EBV-LMP1-positive nasopharyngeal carcinoma(NPC)was determined in our study.We show that EBV-LMP1 exhibits a new function in remodeling mitochondrial morphology by activating Drp1.A high level of p-Drp1(Ser616)or a low level of p-Drp1(Ser637)correlates with poor overall survival and disease-free survival.Furthermore,the protein level of p-Drp1(Ser616)is related to the clinical stage(TNM stage)of NPC.Targeting Drp1 impairs mitochondrial function and induces cell death in LMP1-positive NPC cells.In addition,EBV-LMP1 regulates Drp1 through two oncogenic signaling axes,AMPK and cyclin B1/Cdk1,which promote cell survival and cisplatin resistance in NPC.Our findings provide novel insight into the role of EBV-LMP1-driven mitochondrial fission in regulating Drp1 phosphorylation at serine 616 and serine 637.Disruption of Drp1 could be a promising therapeutic strategy for LMP1-positive NPC.

Let-7i miRNA and platinum loaded nano-graphene oxide platform for detection/reversion of drug resistance and synergetic chemical-photothermal inhibition of cancer cell

The drug resistance of chemotherapy is a major challenge to overcome for antineoplastic agents and the reverse of drug resistant is essential for cancer therapy. Herein, we developed a drug delivery system which can simultaneously detect/reverse the drug resistance and perform synergetic treatment of cancer. In this work, we integrated cyanine5(Cy5) modified mi RNA(let-7 i)(Cy5-mi RNA) and platinum onto nano-graphene oxide(NGO)(30-50 nm) platform to achieve simultaneously detection/reversion of drug resistance and synergetic treatment of cisplatin resistant SKOV3 cells(SKOV3 DDP cells). The Cy5-mi RNA adsorbed on NGO could selectively bind the drug resistance related m RNA follow by suppress the expression of drug resistance m RNA, and the binding simultaneously induced the release of Cy5-mi RNA from the NGO, thus the fluorescence signal of Cy5 recovered and could be used for drug resistance monitoring.Moreover, the mi RNA suppressed the Cyclin D1 protein expressions thus reversed the drug resistance. The loaded platinum(Ⅳ)(Pt(Ⅳ)) was converted to the therapeutic platinum(Ⅱ)(Pt(Ⅱ)) in both tumor acidic and reductive environment responsive behavior. NGO furtherly performed photothermal therapy under near infrared(NIR) laser irradiation and enhanced the therapeutic effect. All in all, this nanoplatform realized detection/reversion of the drug resistance as well as synergetic chemical-photothermal treatment of ovarian cancer cells, which holds great promise in the treatment of drug resistant cancer cells.

Living cell for drug delivery

The living cells have been emerged as useful platforms for drug delivery due to their advantages of good liquidity,stability,and low immunogenicity.In this review,we summarized the development of living cells-based drug delivery systems.The drug loading methods,applications,and advantages of living cell drug delivery systems were summarized.Different living cells for drug delivery,the mechanisms of action,therapeutic applications,as well as main features were summarized and highlighted.The recent research progress and challenges were discussed.The future directions and prospects were proposed.

Targeting the signaling in Epstein-Barr virus-associated diseases: mechanism, regulation, and clinical study

Epstein-Barr virus-associated diseases are important global health concerns.As a group I carcinogen,EBV accounts for 1.5%of human malignances,including both epithelial-and lymphatic-originated tumors.Moreover,EBV plays an etiological and pathogenic role in a number of non-neoplastic diseases,and is even involved in multiple autoimmune diseases(SADs).In this review,we summarize and discuss some recent exciting discoveries in EBV research area,which including DNA methylation alterations,metabolic reprogramming,the changes of mitochondria and ubiquitin-proteasome system(UPS),oxidative stress and EBV lytic reactivation,variations in non-coding RNA(ncRNA),radiochemotherapy and immunotherapy.Understanding and learning from this advancement will further confrm the far-reaching and future value of therapeutic strategies in EBV-associated diseases.