Qingjian Zou

The FKBP-based chemically induced dimerization (CID) technology is a fundamental tool for spatiotemporal precise modulation of protein functions in living cells, widely used in gene editing, protein function regulation, disease therapy, and drug development. However, its widespread application is limited by the inherent drawbacks including issues with immunosuppressive activity, stability, reversibility, and in vivo delivery. Recent years have seen remarkable progress in addressing these challenges: orthogonalization strategies eliminate immunosuppressive effects, fast-dissociating ligands and optogenetic systems enable reversible regulation, and protein engineering optimizes the FKBP/FRB domains to enhance stability and reduce immunogenicity. Moreover, integration with novel delivery technologies broadens its application scope greatly. This review summarizes the key optimization strategies and innovative applications of this technology in the cutting-edge biological research, aiming to reference the development of next-generation chemogenetic tools with higher precision, better safety, and greater application potential.

Parkinson’s disease (PD) has been associated with the microbiota-gut-brain axis, and probiotics have been shown to alleviate PD symptoms via various mechanisms. Akkermansia muciniphila has demonstrated therapeutic effects in some neurological disorders; however, its role in PD treatment remains debated. This study revealed that orally administered hypoactive Akkermansia muciniphila inhibited dopaminergic neuron loss in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse model. Its ingestion also reduced astrocyte activation and mitigated inflammatory responses in both the brain and colon. Additionally, hypoactive Akkermansia muciniphila modulated intestinal microbiota composition in PD model mice. These findings suggest that hypoactive Akkermansia muciniphila reduces neurotoxicity in an MPTP-induced PD model and may offer potential for inhibiting PD progression. IMPORTANCE: Recent research suggests a connection between Parkinson’s disease (PD) and the microbiota-gut-brain axis, with evidence indicating that probiotics may alleviate PD symptoms. Although Akkermansia muciniphila has demonstrated potential benefits for certain neurological disorders, its efficacy in the treatment of PD is still a subject of ongoing debate. Here, we demonstrated that hypoactive Akkermansia muciniphila ameliorated dopaminergic neuronal death, correlating to the reduction of glial hyperactivation and neuroinflammation. Hypoactive Akkermansia muciniphila also induced microbiota fluctuation, which may perform a sophisticated effect on PD progression. Our study may provide an innovative strategy for using hypoactive Akkermansia muciniphila as a therapy or adjuvant therapy for PD.

Messenger RNA (mRNA), emerging as a revolutionary therapeutic tool, has shown remarkable potential in diverse fields such as vaccine development, tumor immunotherapy, gene therapy, and protein replacement therapy, attributed to its high programmability and safety. However, traditional engineered mRNA therapeutics encounter obstacles like short half‐lives and limited protein expression, which impede their extensive application in the biomedical domain. In recent years, with the progress of RNA modification technologies and the advent of novel RNA techniques, including mRNA, self‐amplifying RNA (saRNA), circular RNA (circRNA), and branched RNA (brRNA), researchers have achieved substantial breakthroughs in enhancing RNA stability, prolonging protein expression, and reducing immunogenicity. This article comprehensively reviews the structure, function, optimization strategies, and biomedical applications of these protein‐coding RNAs.

Glycosylase base editor (GBE) can induce C‐to‐G transversion in mammalian cells, showing great promise for the treatment of human genetic disorders. However, the limited efficiency of transversion and the possibility of off‐target effects caused by Cas9 restrict its potential clinical applications. In our recent study, we have successfully developed TaC9‐CBE and TaC9‐ABE by separating nCas9 and deaminase, which eliminates the Cas9‐dependent DNA off‐target effects without compromising editing efficiency. We developed a novel GBE called TaC9‐GBE^YE1, which utilizes the deaminase and UNG‐nCas9 guided by TALE and sgRNA, respectively. TaC9‐GBE^YE1 showed comparable levels of on‐target editing efficiency to traditional GBE at 19 target sites, without any off‐target effects caused by Cas9 or TALE. The TaC9‐GBE^YE1 is a safe tool for gene therapy.

Spinal muscular atrophy (SMA) is a devastating neuromuscular disease caused by mutations in the survival motor neuron 1 (SMN1) gene. Gene editing technology repairs the conversion of the 6th base T to C in exon 7 of the paralogous SMN2 gene, compensating for the SMN protein expression and promoting the survival and function of motor neurons. However, low editing efficiency and unintended off‐target effects limit the application of this technology. Here, we optimized a TaC9‐adenine base editor (ABE) system by combining Cas9 nickase with the transcription activator‐like effector (TALE)‐adenosine deaminase fusion protein to effectively and precisely edit SMN2 without detectable Cas9 dependent off‐target effects in human cell lines. We also generated human SMA‐induced pluripotent stem cells (SMA‐iPSCs) through the mutation of the splice acceptor or deletion of the exon 7 of SMN1. TaC9‐R10 induced 45% SMN2 T6 > C conversion in the SMA‐iPSCs. The SMN2 T6 > C splice‐corrected SMA‐iPSCs were directionally differentiated into motor neurons, exhibiting SMN protein recovery and antiapoptosis ability. Therefore, the TaC9‐ABE system with dual guides from the combination of Cas9 with TALE could be a potential therapeutic strategy for SMA with high efficacy and safety.

Background and Aim: African swine fever (ASF), a globally transmitted viral disease caused by ASF virus (ASFV), can severely damage the global trade economy. Laboratory diagnostic methods, including pathogen and serological detection techniques, are currently used to monitor and control ASF. Because the large double-stranded DNA genome of the mature virus particle is wrapped in a membrane, the stability of ASFV and its genome is maintained in most natural environments. This study aimed to investigate the stability of ASFV under different environmental conditions from both genomic and antibody perspectives, and to provide a theoretical basis for the prevention and elimination of ASFV. Materials and Methods: In this study, we used quantitative real-time polymerase chain reaction for pathogen assays and enzyme-linked immunosorbent assay for serological assays to examine the stability of the ASFV genome and antibody, respectively, under different environmental conditions. Results: The stability of the ASFV genome and antibody under high-temperature conditions depended on the treatment time. In the pH test, the ASFV genome and antibody remained stable in both acidic and alkaline environments. Disinfection tests revealed that the ASFV genome and antibody were susceptible to standard disinfection methods. Conclusion: Collectively, the results demonstrated that the ASFV genome is highly stable in favorable environments but are also susceptible to standard disinfection methods. This study focuses on the stability of the ASFV genome under different conditions and provides various standard disinfection methods for the prevention and control of ASF. Keywords: African swine fever, enzyme-linked immunosorbent assay, quantitative polymerase chain reaction, standard disinfection methods, virus stability.