Efficient Editing of an Adenoviral Vector Genome with CRISPR/Cas9

Immunotherapy based on genetic modification of T cells has played an important role in the treatment of tumors and viral infections. Moreover, adenoviral vectors engineered with improved safety due to their inability to integrate into the host genome have been key in the clinical application of T cell therapy. However, the commonly used adenoviral vector Ad5 exhibits low efficiency of infection of human T cells and the details of the intracellular trafficking pathway of adenoviral vectors in human primary T cells remains unclear. Resolution of these issues will depend on successful modification of the adenoviral vector. To this end, here we describe the successful establishment of a simple and efficient method for editing adenoviral vectors in vitro using the CRISPR-Cas9 gene editing system to target the adenoviral fiber gene. Electronic supplementary materialThe online version of this article (10.1007/s12088-020-00905-3) contains supplementary material, which is available to authorized users.     

Phosphorylation of Novel Interactors of Wild Soybean GsSnRK1 Protein Kinase

Plant Molecular Biology Reporter - Plant SnRK1 kinases have been reported to participate in regulating many aspects of plant biology including resistance to biotic and abiotic stresses. Wild...     

High-content screening of diterpenoids from Isodon species as autophagy modulators and the functional study of their antiviral activities

Cell Biology and Toxicology - Autophagy is a conserved lysosomal degradation process, and abnormal autophagy has been associated with various pathological processes, e.g., neurodegeneration,...     

SETD2 epidermal deficiency promotes cutaneous wound healing via activation of AKT/mTOR Signalling

ObjectivesCutaneous wound healing is one of the major medical problems worldwide. Epigenetic modifiers have been identified as important players in skin development, homeostasis and wound repair. SET domain–containing 2 (SETD2) is the only known histone H3K36 tri‐methylase; however, its role in skin wound healing remains unclear.Materials and MethodsTo elucidate the biological role of SETD2 in wound healing, conditional gene targeting was used to generate epidermis‐specific Setd2‐deficient mice. Wound‐healing experiments were performed on the backs of mice, and injured skin tissues were collected and analysed by haematoxylin and eosin (H&E) and immunohistochemical staining. In vitro, CCK8 and scratch wound‐healing assays were performed on Setd2‐knockdown and Setd2‐overexpression human immortalized keratinocyte cell line (HaCaT). In addition, RNA‐seq and H3K36me3 ChIP‐seq analyses were performed to identify the dysregulated genes modulated by SETD2. Finally, the results were validated in functional rescue experiments using AKT and mTOR inhibitors (MK2206 and rapamycin).ResultsEpidermis‐specific Setd2‐deficient mice were successfully established, and SETD2 deficiency resulted in accelerated re‐epithelialization during cutaneous wound healing by promoting keratinocyte proliferation and migration. Furthermore, the loss of SETD2 enhanced the scratch closure and proliferation of keratinocytes in vitro. Mechanistically, the deletion of Setd2 resulted in the activation of AKT/mTOR signalling pathway, while the pharmacological inhibition of AKT and mTOR with MK2206 and rapamycin, respectively, delayed wound closure.ConclusionsOur results showed that SETD2 loss promoted cutaneous wound healing via the activation of AKT/mTOR signalling.     

Fitness of F1 hybrids between 10 maternal wild soybean populations and transgenic soybean

Transgenic Research - The releasing of transgenic soybeans (Glycine max (L.) Merr.) into farming systems raises concerns that transgenes might escape from the soybeans via pollen into...     

新工科背景下生物反应工程课程的多元化教学模式 改革与探索

生物反应工程作为一门理论性与应用性都很强的专业课程,在生物工程等相关专业的课程设置中处于桥梁和纽带地位,对新型应用型工科人才的培养发挥着重要的作用。但由于该课程中公式等抽象理论知识过多,导致学生学习效率十分低下。因此,为了适应新工科教育背景下对创新型人才培养的需求,提高学生的学习兴趣和积极性,并培养学生的自主学习等创新能力,教学团队在课程教学中通过引入虚拟仿真技术、开展微课教学、采用案例式教学模式、利用科研平台等多元化方式,对该课程的教学模式、方法和手段尝试改革和探索,取得了一定的教学效果,并就此进行了一些探讨,以期能为相关课程的教学改革提供一些思路和启示。     

柚皮苷通过P2X7受体减轻HIV-1包膜糖蛋白gp120导致大鼠神经功能损害的的作用及机制

人类免疫缺陷病毒-1(Human immunodeficiency virus-1,HIV-1)包膜糖蛋白gp120能够引起大鼠出现学习记忆障碍的行为学改变并增加海马中P2X7受体的表达;柚皮苷对gp120引起的行为学改变及P2X7表达增多具有改善作用。为了研究柚皮苷通过减少P2X7受体表达减轻gp120导致大鼠神经功能损害的作用及机制,本实验选择SD大鼠并分为假手术操作的对照组、脑室注射gp120的gp120组、脑室注射gp120及不同剂量柚皮苷灌胃的柚皮苷组、脑室注射BzATP的BzATP组、脑室注射gp120、BzATP及90mg/kg柚皮苷灌胃的90mg/kg/d柚皮苷+BzATP组。Morris水迷宫检测逃避潜伏期及错误次数,TUNEL法检测海马中细胞凋亡率,Elisa法检测海马中肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白介素-1β(Interleukin-1β,IL-1β)、白介素-6(Interleukin-6,IL-6)的含量,Western blot检测海马中P2X7受体的表达。结果显示:与对照组比较,gp120组大鼠的逃避潜伏期延长,错误次数及海马中细胞凋亡率、TNF-α、IL-1β、IL-6的含量、P2X7受体的表达水平增加(P<0.05);与gp120组比较,不同剂量柚皮苷组大鼠的逃避潜伏期缩短,错误次数及海马中细胞凋亡率、TNF-α、IL-1β、IL-6的含量、P2X7受体的表达水平减少(P<0.05);与90mg/kg/d柚皮苷组比较,90mg/kg/d柚皮苷+BzATP组大鼠的逃避潜伏期延长,错误次数及海马中细胞凋亡率、TNF-α、IL-1β、IL-6的含量、P2X7受体的表达水平增加(P<0.05)。本研究的结果表明柚皮苷减轻HIV-1包膜糖蛋白gp120诱导的神经功能损害,这一作用与抑制海马组织中P2X7受体表达并减轻炎症反应及细胞凋亡有关。创新在于首次阐明了柚皮苷减轻HIV-1包膜糖蛋白gp120诱导神经功能损害的分子机制。在gp120引起大鼠行为学改变及海马组织中细胞凋亡、炎症反应激活的过程中,柚皮苷通过抑制P2X7受体的表达来改善行为学改变、抑制细胞凋亡及炎症反应的激活。