Delivery of CRISPR/Cas9 for therapeutic genome editing

The clustered, regularly‐interspaced, short palindromic repeat (CRISPR)‐associated nuclease 9 (CRISPR/Cas9) is emerging as a promising genome‐editing tool for treating diseases in a precise way, and has been applied to a wide range of research in the areas of biology, genetics, and medicine. Delivery of therapeutic genome‐editing agents provides a promising platform for the treatment of genetic disorders. Although viral vectors are widely used to deliver CRISPR/Cas9 elements with high efficiency, they suffer from several drawbacks, such as mutagenesis, immunogenicity, and off‐target effects. Recently, non‐viral vectors have emerged as another class of delivery carriers in terms of their safety, simplicity, and flexibility. In this review, we discuss the modes of CRISPR/Cas9 delivery, the barriers to the delivery process and the application of CRISPR/Cas9 system for the treatment of genetic disorders. We also highlight several representative types of non‐viral vectors, including polymers, liposomes, cell‐penetrating peptides, and other synthetic vectors, for the therapeutic delivery of CRISPR/Cas9 system. The applications of CRISPR/Cas9 in treating genetic disorders mediated by the non‐viral vectors are also discussed.     

Determination of ionophore antibiotics nactins produced by fecal Streptomyces from sheep

To investigate the correlation between fecal actinobacteria and host animals, Streptomyces was isolated from fresh faeces of healthy sheep and secondary metabolites were analyzed. The most frequently isolated strain S161 with antibiotic activity against bacteria and fungi were analyzed. The S161 showed the highest 99 % similarity to Streptomyces canus DSB17 based on the 16S rRNA gene sequence analysis. Metabolite analysis based on MS and NMR spectra showed that S161 produces nactins, cyclotetralactones derived from nonactic acid and homononactic acid as building units of ionophoretic character. Due to ionophores are antimicrobial compounds that are commonly fed to ruminant animals to improve feed efficiency, stable beneficial interactions between Streptomyces bacteria and vertebrates have been demonstrated.     

Isolation and characterization of endophytic streptomycete antagonists of Fusarium wilt pathogen from surface-sterilized banana roots

A total of 131 endophytic actinomycete strains were successfully isolated from surface-sterilized banana roots. These isolates belonged to Streptomyces (n=99), Streptoverticillium (n=28), and Streptosporangium (n=2) spp. The remaining 2 isolates were not identified. About 18.3% of the isolates inhibited the growth of pathogenic Fusarium oxysporum f. sp. cubense on banana tissue extract medium. The most frequently isolated Streptomyces sp. strain S96 was similar to Streptomyces griseorubiginosus. About 37.5% of the S. griseorubiginosus strains were antagonistic to F. oxysporum f. sp. cubense. The antagonism of strain S96 was lost when FeCl(3) was introduced into the inhibition zone. In vivo biocontrol assays showed that the disease severity index (DSI) was significantly (P=0.05) reduced and mean fresh weight increased (P=0.001) in plantlets treated with strain S96 compared to those grown in the absence of the biocontrol strain. These findings indicate the potential of developing siderophore-producing Streptomyces endophytes for the biological control of fusarium wilt disease of banana.     

Fungal diversity in sheep (Ovis aries) and cattle (Bos taurus) feces assessed by comparison of 18S, 28S and ITS ribosomal regions

To explore the fungal diversity in ruminant feces for bioenergy, libraries based on internal transcribed spacer (ITS), 18S rRNA, and 28S rRNA regions were constructed, respectively. Although the libraries were constructed from the same DNA extracts, the fungal taxa analyses based on these libraries are different. The ITS and 28S libraries comprised higher proportions of fungal clones than 18S libraries, and the ITS libraries converged into the lower diversities. The ITS libraries could be used to analyze the fungal community. The 18S libraries were suitable for the fungi and protozoa community. However, the 28S are suitable for analysis of Ascomycota fungi. The major fungal taxa in cattle feces analyzed by ITS, 18S, and 28S libraries are similar to those of sheep feces, respectively. The fungal taxa detected by the ITS library comprised only 20 % fungal taxa detected by the three libraries. The 18S library comprised 30 % fungal taxa; the 28S library comprised about 50 % fungal taxa. The results indicated that primer sets toward different DNA regions lead to the difference in structures of fungal community. So the selection of primer sets may influence the fungal communities, and libraries based on single primer sets may underestimate the fungal diversity. The comparison of ITS, 18S, and 28S libraries could fid more diverse fungi than that based on only one library.     

初夏渤海湾初级生产力分布特征及影响因素

根据2016年初夏在渤海湾周边重要经济开发区外海域所进行的调查和研究数据进行分析,利用稳定同位素¹³C示踪技术估算海域内初级生产力（Primary Productivity,PP）等,同时结合叶绿素a（Chlorophyll-a,Chl a）,营养盐以及透明度等水文环境参数,深入分析和探讨初夏渤海湾环境特征对浮游植物生物量（Chl a）和初级生产力的影响。结果表明：受陆地径流和渤海中部冷流输入等因素的影响,调查海域呈现3个温盐特征差异显著的海区,即近岸高温低盐海区、中部高温高盐海区和湾口低温高盐海区。Chl a受温度和营养盐等因素影响整体呈现近岸高湾口低、表层高底层低的分布特征,含量变化范围为1.27-20.82 mg/m³;在近岸营养盐含量充足,温度适宜,浮游植物生产旺盛,Chl a平均含量达（8.37±2.90）mg/m³,其中表层近27.5%水样中含量超10 mg/m³,存在发生赤潮的风险。而在中部和湾口区域受营养盐限制和温度的影响,Chl a含量远低于近岸。PP整体水平在44.79-792.73 mg C m^-3 d^-1之间,平均为（144.13±137.79）mg C m^-3 d^-1。其中近岸营养盐含量充足浮游植物生长旺盛,初级生产力水平较高,而中部和湾口海域受营养盐和温度的限制,初级生产力水平较低。初级生产力指数I（同化系数）变化范围在0.79-5.90 mg C/（mg Chl a·h）之间,平均为（3.40±1.33）mg C/（mg Chl a·h）。利用标准深度积分模型对水柱初级生产力（Depth-integrated primary productivity,∑PP）进行估算,结果表明其范围在56.88-772.31 mg C m^-2 d^-1之间,平均为（232.26±126.47）mg C m^-2 d^-1,近岸受陆源输入影响Chl a较高,在天津海河口和黄骅市排污河外出现高值点,受透明度的影响,中部和湾口部分站点出现高生产力区。     

Single-cell transcriptomics of cardiac progenitors reveals functional subpopulations and their cooperative crosstalk in cardiac repair

Dear Editor,Myocardial infarction is one of the leading causes of morbidity and mortality.Stem/progenitor cells therapy has emerged as a promising strategy for the cardiac repair,especially those derived from cardiac tissue,have attracted worldwide attention(Tompkins et al.,2018).However,challenges and controversies remain in characterizing functional progenitors and explaining their mechanisms of action.     

Nitrogen removal from synthetic wastewater using single and mixed culture systems of denitrifying fungi,bacteria, and actinobacteria

Applied Microbiology and Biotechnology - The aim of this study was to investigate the effects of single and mixed culture of denitrifying fungi, bacteria, and actinobacteria on nitrogen removal and...