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.

Multiple internal resonances of rotating composite cylindrical shells under varying temperature fields

Composite cylindrical shells,as key components,are widely employed in large rotating machines.However,due to the frequency bifurcations and dense frequency spectra caused by rotation,the nonlinear vibration usually has the behavior of complex multiple internal resonances.In addition,the varying temperature fields make the responses of the system further difficult to obtain.Therefore,the multiple internal resonances of composite cylindrical shells with porosities induced by rotation with varying temperature fields are studied in this paper.Three different types of the temperature fields,the Coriolis forces,and the centrifugal force are considered here.The Hamilton principle and the modified Donnell nonlinear shell theory are used to obtain the equilibrium equations of the system,which are transformed into the ordinary differential equations(ODEs)by the multi-mode Galerkin technique.Thereafter,the pseudo-arclength continuation method,which can identify the regions of instability,is introduced to obtain the numerical results.The detailed parametric analysis of the rotating composite shells is performed.Multiple internal resonances caused by the interaction between backward and forward wave modes and the energy transfer phenomenon are detected.Besides,the nonlinear amplitude-frequency response curves are different under different temperature fields.

VEGF-B prevents excessive angiogenesis by inhibiting FGF2/FGFR1 pathway

Although VEGF-B was discovered as a VEGF-A homolog a long time ago,the angiogenic effect of VEGF-B remains poorly understood with limited and diverse findings from different groups.Notwithstanding,drugs that inhibit VEGF-B together with other VEGF family members are being used to treat patients with various neovascular diseases.It is therefore critical to have a better understanding of the angiogenic effect of VEGF-B and the underlying mechanisms.Using comprehensive in vitro and in vivo methods and models,we reveal here for the first time an unexpected and surprising function of VEGF-B as an endogenous inhibitor of angiogenesis by inhibiting the FGF2/FGFR1 pathway when the latter is abundantly expressed.Mechanistically,we unveil that VEGF-B binds to FGFR1,induces FGFR1/VEGFR1 complex formation,and suppresses FGF2-induced Erk activation,and inhibits FGF2-driven angiogenesis and tumor growth.Our work uncovers a previously unrecognized novel function of VEGF-B in tethering the FGF2/FGFR1 pathway.Given the anti-angiogenic nature of VEGF-B under conditions of high FGF2/FGFR1 levels,caution is warranted when modulating VEGF-B activity to treat neovascular diseases.

An Attenuated Highly Pathogenic Chinese PRRS Viral Vaccine Confers Cross Protection to Pigs against Challenge with the Emerging PRRSV NADC30-Like Strain

A novel PRRSV strain was isolated in China that was genetically similar to the NADC30 strain which is reported to have spread throughout China. The objective of the present study was to evaluate the cross-protective efficacy of the live vaccine TJM-F92 in young pigs against challenge with a NADC30-like strain, HN201605. Twenty-five PRRSV-and antibody-free pigs were randomly divided into the following five groups: Vac/ChA, Unvac/ChA, Vac/ChB, Unvac/ChB and the mock.The pigs in groups Vac/ChA and Vac/ChB were inoculated intramuscularly with 1 mL TJM-F92(10^(5.0) TCID_(50)/mL). At28 days post vaccination(0 days post challenge), groups Vac/ChA and Unvac/ChA were inoculated intranasally with 10^(4.5) TCID_(50)/mL PRRSV strain TJ F3(2 mL/pig), while groups Vac/ChB and Unvac/Ch B were inoculated, using the same route, with the same dose of the NADC30-like strain HN201605 F3. Protective effects of the PRRSV strain were observed in all pigs in the Vac/ChA and Vac/ChB groups. Neither high fever nor signs of clinical disease were observed through the experiment in these groups, whereas pigs in Unvac/ChA group exhibited serious clinical symptoms, pathological lesions,and weight loss. In Unvac/ChB group, pigs developed milder clinical symptoms, which demonstrated that the NADC30-like strain HN201605 had moderate pathogenicity. The results suggest that the MLV vaccine strain TJM-F92 is an effective and safe vaccine candidate for use in China.

蝙蝠中菊头ACE2蛋白和多个动物ACE2是蝙蝠病毒RaTG13和新冠病毒的功能受体

新冠病毒可能起源于蝙蝠,但是否存在中间宿主以及可能的中间宿主是什么,目前还没有明确答案.来自于中菊头蝙蝠的冠状病毒RaT G13是所有已知冠状病毒中与新冠病毒基因组同源性最高的,被认为可能是新冠病毒的起源.本研究首次报道中菊头angiotensin-converting enzyme 2(ACE2)不仅能介导RaT G13入侵,也能介导新冠病毒入侵,但新冠病毒受体结合区(Receptor binding domain,RBD)与中菊头ACE2的亲和力明显比人ACE2低.随后,本研究评估了来自9个目16个不同物种的ACE2作为RaT G13、新冠病毒以及SARS病毒的入侵受体的可能性,发现7个物种的ACE2能介导所有三种病毒的入侵,提示这三个病毒可能具有较广的宿主范围.另外,本研究发现RaT G13和新冠病毒S蛋白在结合小鼠和穿山甲ACE2时存在明显差异.进一步研究发现病毒S蛋白的第484和498位氨基酸对识别小鼠ACE2至关重要.该研究对深入了解冠状病毒入侵、宿主范围以及病毒宿主共进化的机制具有积极意义.