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- Keywords：巢裸胞壳菌;海洋真菌;芳香聚酮;多硫代二酮哌嗪;细胞毒6|22|0
Updated：2023-08-17 - Abstract：目的鉴定褐飞虱中的piggyBac类似元素并评估其转座活性。创新点 首次鉴定了褐飞虱中的piggyBac类似元素并发现NlPLE25具备转座活性。方法使用RepeatMasker和RepeatModeler软件预测褐飞虱基因组中的piggyBac类似元素序列。随后在S2细胞中测试了NlPLE25的切割活性，在Sf9细胞中测试了NlPLE25的转座能力。结论我们从褐飞虱基因组中鉴定出28个piggyBac类似元素。进一步分析NlPLE25发现其在昆虫细胞中引起精确的切除和转座。此外，我们还发现了NlPLE25与piggyBac转座子之间的交叉识别。Keywords：褐飞虱;piggyBac类似元素;转座子;NlPlE2537|10|0Updated：2022-05-30
- Abstract：The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)-reporter system to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAMs of 5'-NCC-3' and 5'-NTC-3' are not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.Keywords：CRISPR/Cas;SpRY;Protospacer adjacent motif (PAM);Recognize32|8|0Updated：2022-05-30
- Abstract：Ulcerative colitis (UC) is a chronic and recurrent inflammatory bowel disease (IBD) that has become a major gastroenterologic problem during recent decades. Numerous complicating factors are involved in UC development such as oxidative stress, inflammation, and microbiota disorder. These factors exacerbate damage to the intestinal mucosal barrier. Spirulina platensis is a commercial alga with various biological activity that is widely used as a functional ingredient in food and beverage products. However, there have been few studies on the treatment of UC using S. platensis aqueous extracts (SP), and the underlying mechanism of action of SP against UC has not yet been elucidated. Herein, we aimed to investigate the modulatory effect of SP on microbiota disorders in UC mice and clarify the underlying mechanisms by which it alleviates damage to the intestinal mucosal barrier. Dextran sulfate sodium (DSS) was used to establish a normal human colonic epithelial cell (NCM460) injury model and UC animal model. The mitochondrial membrane potential assay 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and staining with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) and Hoechst 33258 were carried out to determine the effects of SP on the NCM460 cell injury model. Moreover, hematoxylin and eosin (H&E) staining, transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qPCR), western blot, and 16S ribosomal DNA (rDNA) sequencing were used to explore the effects and underlying mechanisms of action of SP on UC in C57BL/6 mice. In vitro studies showed that SP alleviated DSS-induced NCM460 cell injury. SP also significantly reduced the excessive generation of intracellular reactive oxygen species (ROS) and prevented mitochondrial membrane potential reduction after DSS challenge. In vivo studies indicated that SP administration could alleviate the severity of DSS-induced colonic mucosal damage compared with the control group. Inhibition of inflammation and oxidative stress was associated with increases in the activity of antioxidant enzymes and the expression of tight junction proteins (TJs) post-SP treatment. SP improved gut microbiota disorder mainly by increasing antioxidant enzyme activity and the expression of TJs in the colon. Our findings demonstrate that the protective effect of SP against UC is based on its inhibition of pro-inflammatory cytokine overproduction, inhibition of DSS-induced ROS production, and enhanced expression of antioxidant enzymes and TJs in the colonic mucosal barrier.Keywords：Spirulina platensis aqueous extracts;Ulcerative colitis;Oxidative stress;Inflammation;Antioxidant;Gut microbiota85|16|0Updated：2022-05-30
- Abstract：The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)-reporter system to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAMs of 5'-NCC-3' and 5'-NTC-3' are not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.Keywords：CRISPR/Cas;SpRY;Protospacer adjacent motif (PAM);Recognize245|2|0Updated：2022-05-30
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