JOURNAL OF EXPERIMENTAL BOTANY
·
2020
·Vol. 71
·期 19
·5948-5962
·
IF 5.908
Deng Changping,Shi Min,Fu Rong,Zhang Yi,Wang Qiang,Zhou Yang,Wang Yao,Ma Xingyuan,Kai Guoyin
Phenolic acids and tanshinones are major bioactive ingredients in Salvia miltiorrhiza, which possess pharmacological activities with great market demand. However, transcriptional regulation of phenolic acid and tanshinone biosynthesis remains poorly understood. Here, a basic leucine zipper transcription factor (TF) named SmbZIP1 was screened from the abscisic acid (ABA)-induced transcriptome library. Overexpression of SmbZIP1 positively promoted phenolic acid biosynthesis by enhancing expression of biosynthetic genes such as cinnamate-4-hydroxylase (C4H1). Furthermore, biochemical experiments revealed that SmbZIP1 bound the G-Box-like1 element in the promoter of the C4H1 gene. Meanwhile, SmbZIP1 inhibited accumulation of tanshinones mainly by suppressing the expression of biosynthetic genes including geranylgeranyl diphosphate synthase (GGPPS) which was confirmed as a target gene by in vitro and in vivo experiments. In contrast, the phenolic acid content was reduced and tanshinone was enhanced in CRISPR/Cas9 [clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9]-mediated knockout lines. In addition, the previously reported positive regulator of tanshinone biosynthesis, SmERF1L1, was found to be inhibited in SmbZIP1 overexpression lines indicated by RNA sequencing, and was proven to be the target of SmbZIP1. In summary, this work uncovers a novel regulator and deepens our understanding of the transcriptional and regulatory mechanisms of phenolic acid and tanshinone biosynthesis, and also sheds new light on metabolic engineering in S. miltiorrhiza.
研究领域:
植物生物学
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分子生物学
·
药理学
·
基因工程
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DOI
PubMed
高影响因子
Cell Reports Medicine
·
2026
·
IF 14
Chen Zhang,Tianyu Li,Huoye Hao,Gang Wang,Yuzhe Zhang,Yue Huang,Lili Chen,Jin Wang,Xiang-Jiao Yang,Xiuli Bi
The primary pathogenic mechanism of inflammatory bowel disease (IBD) involves elevated levels of reactive oxide species (ROS) in the gut, leading to oxidative stress and damage to the intestinal barrier function. We engineered a non-pathogenic bacterial strain, Escherichia coli Nissle 1917 (EcN), for oral CRISPR-Cas9 delivery to edit NOX2 (encoding the gp91 phox subunit of NADPH oxidase 2), thereby alleviating IBD symptoms by reducing ROS. EcN expressing the Cas9/sgRNA ribonucleoprotein (RNP) was encapsulated in a hydrogel (composed of hyaluronic acid, chitosan, and MgCl 2 ), which protected EcN-RNP from degradation and increased survival from 0.07% to 12%. EcN-RNP hydrogel reduced NOX2 expression by 47% and ROS by 88% in lipopolysaccharide-induced RAW264.7 cells. Moreover, oral delivery of EcN-RNP hydrogel mitigated inflammation in dextran sodium sulfate-induced colitis mouse models. Mechanistically, the hydrogel could activate the NRF2-HO1/GPX4 pathway by inhibiting NOX2 expression, enhancing oxidative defense, and promoting glutathione accumulation. Thus, the EcN-RNP hydrogel offers a promising therapeutic strategy for IBD.
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PubMed
高影响因子
Lipids in Health and Disease
·
2021
·Vol. 20
·期 1
·1-11
·
IF 3.876
Zha Yiwen,Lu Yaoyao,Zhang Ting,Yan Kunning,Zhuang Wenwen,Liang Jingyan,Cheng Yong,Wang Yingge
Background High levels of apolipoprotein C3 (APOC3) can lead to hypertriglyceridemia, which increases the risk of cardiovascular disease. We aim to create APOC3-knockout (KO) rabbits and explore the effects of APOC3 deletion on the occurrence and development of atherosclerosis.Methods An sgRNA anchored to exon 2 of APOC3 was designed to edit embryo genomes using the CRISPR/Cas9 system. The founder rabbits were sequenced, and their lipid profile, inflammatory cytokines, and atherosclerotic plaques were analyzed.Results When given a normal chow (NC) diet, all APOC3-KO rabbits had 50% lower triglyceride (TG) levels than those of the matched age control group. Additionally, their plasma lipoprotein lipase increased. When fed a high-fat diet, APOC3 deficiency was observed to be more conducive to the maintenance of plasma TG, total cholesterol, and low-density lipoprotein cholesterol levels, and the inhibition of the inflammatory response and the protection against atherosclerosis in rabbits.Conclusion APOC3 deficiency can delay the formation of atherosclerosis-induced HFD in rabbits, indicating this is a novel therapeutic target to treat atherosclerosis.
研究领域:
分子生物学
·
心血管疾病
·
基因工程
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DOI
PubMed
高影响因子
Scientific Reports
·
2017
·Vol. 7
·期 1
·1-8
·
IF 4.259
Guo Dongsheng,Liu Haikun,Ruzi Aynisahan,Gao Ge,Nasir Abbas,Liu Yanli,Yang Fan,Wu Feima,Xu Guosheng,Li Yin-xiong
Congenital hyperinsulinism (CHI) is a rare genetic disorder characterized by excess insulin secretion, which results in hypoglycemia. Mutation of sulfonylurea receptor 1 (SUR1), encoded by the ABCC8 gene, is the main cause of CHI. Here, we captured the phenotype of excess insulin secretion through pancreatic differentiation of ABCC8 -deficient stem cells generated by the CRISPR/Cas9 system. ABCC8- deficient insulin-producing cells secreted higher insulin than their wild-type counterparts, and the excess insulin secretion was rescued by nifedipine, octreotide and nicorandil. Further, we tested the role of SUR1 in response to different potassium levels and found that dysfunction of SUR1 decreased the insulin secretion rate in low and high potassium environments. Hence, pancreatic differentiation of ABCC8- deficient cells recapitulated the CHI disease phenotype in vitro , which represents an attractive model to further elucidate the function of SUR1 and to develop and screen for novel therapeutic drugs.
研究领域:
遗传学
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干细胞研究
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分子生物学
·
内分泌学
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DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Liansheng Fan,Suchen Qu,Jing Qian,Gaoyu Shi,Qiqing Huang,Yanni Song,Yu Ji,Quli Fan,Xin Han
Immune checkpoint blockade (ICB) has shown significant progress in treating triple negative breast cancer (TNBC). However, the standalone effectiveness of ICB is modest, offering patients only slight benefits. In this report, we utilize a hypoxia-responsive duplex genome editing strategy to target tumor cell programmed cell death ligand 1 (PD-L1) and protein tyrosine phosphatase non-receptor type 2 (PTPN2), which is synergized with photothermal therapy (PTT) to amplify the therapeutic effect of TNBC. In our approach, the melanin nanoparticles (MNP) serve as the delivery platform for transporting the CRISPR-Cas PTPN2/PD-L1 (Cas-TD) system, which is cross-linked by a hypoxic-sensitive Azo linker and then further modified with tumor homing phenylboronic acid (PBA), resulting in a stimuli-responsive multifunctionality nanotheranostic platform (MPA@Cas-TD@PP). Aided by the guidance of PBA, nanoparticles selectively accumulate at tumor and then disintegrate with enhanced intracellular penetration. Importantly, PTT initiates the immunogenic cell death (ICD), turning the “cold” tumor into a “hot” one, laying the foundation for subsequent ICB. Inhibiting PTPN2 and PD-L1 can synergistically sensitize tumors to immunotherapy, which exhibit magnified immunotherapy effects by increasing activated cytotoxic CD8 + T cells and enhancing interferon-γ (IFN-γ)-mediated effects. In summary, this multifunctional nanotherapeutic platform provides a promising treatment approach for synergistic phototherapy and immunotherapy in TNBC.
研究领域:
肿瘤学
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基因编辑
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纳米医学
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免疫治疗
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产品关联
DOI
高影响因子
Advanced Healthcare Materials
·
2025
·
IF 9.6
Liuxian Chen,Qin Huang,Yongjie Liu,Ke Chen,Qing Yang,Hua Tang,Dan Wang,Zhenrong Tang
Complete hepatitis B virus (HBV) cure is hindered primarily by the stable persistence of covalently closed circular DNA (cccDNA). Gene editing approaches to eradicate HBV by targeting cccDNA face challenges and limitations due to suboptimal editing efficiency and substantial off-target effects. Herein, a combinatorial therapeutic strategy is developed that integrates CRISPR/Cas9-mediated cccDNA disruption with an antisense oligonucleotide (ASO)-targeted degradation of pregenomic RNA (pgRNA). To overcome delivery challenges, a hepatocyte-targeting nanocarrier (UACPG) is engineered, featuring low immunogenicity, high payload capacity, and dual-stimuli responsiveness. The UACPG platform enabled liver-specific delivery through surface-conjugated targeting ligands, followed by on-demand release of Cas9 ribonucleoprotein complexes and ASO via RNase H-dependent degradation and near-infrared (NIR) light activation. The results demonstrated that UACPG can effectively reduce HBV replication and viral antigen levels, while significantly lowering cccDNA in hydrodynamic HBV-infected mouse models, with no significant off-target effects observed. This nanocarrier achieved the spatiotemporally controlled release of gene-editing systems in vitro and in vivo, significantly inhibiting the replication of HBV, thereby establishing an innovative technological platform for developing curative HBV therapies.
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PubMed
高影响因子
Processes
·
2026
·Vol. 14
·期 9
·1347
·
IF 3.4
Zhuoling Ding,Kadireya Tuerxun,Xueqing Luo,Shishui Zhou
γ-Aminobutyric acid (GABA), a major inhibitory neurotransmitter, is known for its physiological functions in alleviating anxiety and improving sleep. Currently, high-yielding GABA food products are mainly obtained through screening wild-type high-producing strains (e.g.,Saccharomyces cerevisiaeisolated from Sichuan pickles yielding 0.67 g/L) or employing co-culture systems (e.g.,Enterococcus faeciumandLactiplantibacillus plantarumreaching 6.35 g/L). While effective, these methods often rely on natural screening strains or multi-microbial interactions. This study employed CRISPR-Cas9 technology to knockout theUGA1gene inSaccharomyces cerevisiae, a key gene responsible for GABA degradation. Starting from the low higher alcoholSaccharomyces cerevisiaeSY-LH, we successfully constructed the recombinant strain SY-LHU. Remarkably, this study discovered a significant upregulation ofGAD1gene expression followingUGA1knockout, which further enhanced GABA synthesis capacity. Under optimal fermentation conditions (inoculum size 4 × 107cells/mL, wort concentration 10 °P, sugar addition 60 g/L, 30 °C for 10 days, and mixing the malt broth every 48 h), the validation fermentation was performed and the GABA content in the wort beverage reached 280.36 mg/L, representing a 385.4% increase compared to the pre-optimization level. Furthermore, sensory evaluation by a trained panel yielded a mean score of 88, with no significant off-flavors detected, demonstrating the product’s high consumer acceptance. This pioneering work provides a novel and feasible technical pathway for developing functional alcoholic beverages with sleep-aiding properties.
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DOI
高影响因子
INDUSTRIAL CROPS AND PRODUCTS
·
2021
·
IF 5.645
Min Shi,Zhiyan Du,Qiang Hua,Guoyin Kai
Phenolic acids comprise the major bioactive compounds in Salvia miltiorrhiza , which can be increased by abscisic acid (ABA) elicitation. However, the underlying regulation mechanism is poorly understood. Here, the ABA-induced transcriptome revealed that bZIP2 was significantly induced, which expressed highly in roots. Bioinformatic analysis revealed that bZIP2 belong to the subgroup A and contained a bZIP domain together with four conservative regions (C1, C2, C3, and C4). Phenolic acid content was increased in bZIP2 knock-out lines medicated by CRISPR/Cas9 technology. In contrast, overexpressing lines produced less phenolic acids by downregulating the PAL gene. Biochemistry assays showed that bZIP2 binds to the ABRE2 element in the promoter region of the PAL gene and suppresses its expression. Besides, bZIP2 physically interacted with SnRK2.3, SnRK2.4, SnRK2.6, and SnRK2.10. Overall, these results demonstrated that bZIP2 is a negative regulator in phenolic acid biosynthesis, which provides new insights for the metabolic engineering of phenolic acids.
研究领域:
植物学
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遗传学
·
代谢工程
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DOI
高影响因子
VIROLOGY
·
2023
·
IF 3.7
Xiaoping Huang,Xiao Wang,Yaqi Sun,Ling Li,Anna Li,Wentao Xu,Xiaolan Xie,Yong Diao
Because it is safe and has a simple genome, recombinant adeno-associated virus (rAAV) is an extremely appealing vector for delivery in in vivo gene therapy. However, its low transduction efficiency for some cells, limits its further application in the field of gene therapy. Bleomycin is a chemotherapeutic agent approved by the FDA whose effect on rAAV transduction has not been studied. In this study, we systematically investigated the effect of Bleomycin on the second-strand synthesis and used CRISPR/CAS9 and RNAi methods to understand the effects of Bleomycin on rAAV vector transduction, particularly the effect of DNA repair enzymes. The results showed that Bleomycin could promote rAAV2 transduction both in vivo and in vitro. Increased transduction was discovered to be a direct result of decreased cytoplasmic rAAV particle degradation and increased second-strand synthesis. TDP1 , PNKP , and SETMAR are required to repair the DNA damage gap caused by Bleomycin, TDP1, PNKP, and SETMAR promote rAAV second-strand synthesis. Bleomycin induced DNA-PKcs phosphorylation and phosphorylated DNA-PKcs and Artemis promoted second-strand synthesis. The current study identifies an effective method for increasing the capability and scope of in-vivo and in-vitro rAAV applications, which can amplify cell transduction at Bleomycin concentrations. It also supplies information on combining tumor gene therapy with chemotherapy.
研究领域:
分子生物学
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基因治疗
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癌症研究
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药理学
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DOI
PubMed
高影响因子
VETERINARY RESEARCH
·
2025
·Vol. 56
·期 1
·1-14
·
IF 3.5
Li Peiyao,Su Rina,Ren Ganglin,Si Hongbin,Song Xingju,Hu Dandan
The apicomplexan parasite Eimeria is the causative agent of coccidiosis, which results in enteritis in animals and imposes a significant economic burden on the farming industry. Apicomplexan Apetala2/ERF (ApiAP2) transcription factors are known to play crucial roles in various processes of Apicomplexa parasites. However, little is understood regarding the role of ApiAP2s in Eimeria species. In this study, we performed a functional characterisation of ApiAP2 through CRISPR-Cas9-mediated knockout. Our results revealed that the parasite’s invasion efficiency, total oocyst output, and virulence were significantly impaired after EtAP2-S1 depletion. RNA sequencing (RNA-Seq) and Cleavage Under Targets and Tagmentation (CUT&Tag) analyses showed that EtAP2-S1 targets the promoters of numerous genes, and its knockout resulted in the up-regulation of 59 sag genes. Furthermore, the knockout strain exhibited significantly lower virulence while offering excellent immune protection, making it a promising candidate for a vaccine. This study demonstrates that EtAP2-S1 is a fitness-conferring gene that suppresses the expression of sag genes in E. tenella , which is the first step in developing gene-knockout vaccines to control coccidiosis.
研究领域:
分子生物学
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寄生虫学
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免疫学
·
兽医学
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产品关联
DOI
PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
·
2019
·Vol. 47
·期 12
·1425-1432
·
IF 3.354
Hongqiu Zhou,Li Yang,Changsuo Wang,Zhiqiang Li,Zhen Ouyang,Mangting Shan,Jun Gu,Yuan Wei
Rat CYP2D1 has been considered as an ortholog of human CYP2D6. To assess the role of CYP2D1 in physiologic processes and drug metabolism, a CYP2D1-null rat model was generated with a CRISPR/Cas9 method. Seven base pairs were deleted from exon 4 of CYP2D1 of Sprague-Dawley wild-type (WT) rats. The CYP2D1-null rats were viable and showed no abnormalities in general appearance and behavior. The metabolism of venlafaxine (VLF) was further studied in CYP2D1-null rats. The Vmax and intrinsic clearance of the liver microsomes in vitro from CYP2D1-null rats were decreased (by ∼46% and ∼57% in males and ∼47% and ∼58% in females, respectively), while the Michaelis constant was increased (by ∼24% in males and ∼25% in females) compared with WT rats. In the pharmacokinetic studies, compared with WT rats, VLF in CYP2D1-null rats had significantly lower apparent total clearance and apparent volume of distribution (decreased by ∼36% and ∼48% in males and ∼23% and ∼25% in females, respectively), significantly increased area under the curve (AUC) from the time of administration to the last time point, AUC from the start of administration to the theoretical extrapolation, and Cmax (increased by ∼64%, ∼59%, and ∼26% in males and ∼43%, ∼35%, and ∼15% in females, respectively). In addition, O-desmethyl venlafaxine formation was reduced as well in CYP2D1-null rats compared with that in WT rats. Rat depression models were developed with CYP2D1-null and WT rats by feeding them separately and exposing them to chronic mild stimulation. VLF showed better efficacy in the WT depression rats compared with that in the CYP2D1-null rats. In conclusion, a CYP2D1-null rat model was successfully generated, and CYP2D1 was found to play a certain role in the metabolism and efficacy of venlafaxine.
研究领域:
药理学
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遗传学
·
分子生物学
·
行为科学
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产品关联
DOI
PubMed
高影响因子
Materials Today Bio
·
2025
·
IF 10.2
Yichen Liu,Dahai Hou,Hui Wang,Qiuying Yan,Yunzhu Xu,Qibiao Wu,Dan Dong,Weixing Shen,Weiwei Tao,Haibo Cheng,Dongdong Sun
Colorectal cancer, a highly invasive malignancy, poses significant treatment challenges due to the immunosuppressive tumor microenvironment. Tumor-associated macrophages, which primarily exhibit the M2 phenotype, play a key role in cancer progression and immune evasion. Reprogramming these M2 macrophages to the tumor-suppressive M1 phenotype represents a promising therapeutic approach to enhance antitumor immunity. In this study, we developed a calcium lactate nanoparticle-based delivery system (CLBRP) to codeliver bufalin, a potent Na + /K + -ATPase inhibitor with anticancer properties, and CRISPR-Cas9, which targets the immune checkpoint CD47. The system releases its payload under low pH conditions typical of the tumor microenvironment, increasing extracellular Ca 2+ and intracellular osmotic pressure, thereby inducing pyroptosis and immunogenic cell death. Bufalin induces cancer cell apoptosis, inhibits Na + /K + -ATPase, and triggers pyroptosis, which activates a strong immune response. Additionally, bufalin and D-lactic acid synergistically promote macrophage repolarization to the M1 phenotype. CRISPR-Cas9-mediated editing of the CD47 gene on tumor cells blocks antiphagocytic signals, enhancing M1 macrophage phagocytosis and increasing the antitumor immune response. This multimodal strategy triggers strong local antitumor effects and controls metastatic lesions via systemic immune activation, demonstrating significant potential for colorectal cancer treatment. Furthermore, this approach offers a new direction for overcoming resistance to immunotherapies and improving clinical outcomes.
研究领域:
肿瘤学
·
免疫治疗
·
纳米医学
·
基因编辑
·
癌症生物学
·
肿瘤微环境
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）