INDUSTRIAL CROPS AND PRODUCTS
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2021
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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
MICROCHEMICAL JOURNAL
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2026
·
IF 5.2
Shuo Zhang,Ting Yang,Rui Lu,Huarong Pang,Huakang Li,Bin Li,Yuzhe Song
Cb Ago selectively depletes wild-type TP53, eliminating background interference for mutant analysis. • Cas9 nickase generates ssDNA initiators to trigger EXPAR under isothermal conditions. • Amplified products disrupt G-quadruplex/ThT fluorescence, enabling a signal-off readout. • The assay achieves 0.14 fM detection limit and single-nucleotide resolution. • Validated in serum and cells; consistent with PCR; discriminates cancer from normal cells. Accurate detection of rare mutant TP53 in the presence of abundant wild-type background remains a major challenge for early diagnosis of colorectal cancer. Here, we report a background-reduced analytical strategy that integrates Cb Ago-mediated wild-type depletion with Cas9 nickase (Cas9n)/sgRNA-triggered exponential amplification reaction (EXPAR) for highly sensitive and specific mutant TP53 analysis. Cb Ago programmed with a guide DNA selectively cleaves wild-type TP53 while preserving the mutant target, thereby eliminating interference from the overwhelming wild-type background. The remaining mutant TP53 is then recognized by two Cas9n/sgRNA complexes, which generate single-stranded initiator fragments through polymerase-assisted strand displacement. These initiators trigger an EXPAR cascade that ultimately disrupts the G-quadruplex structure of a thioflavin T (ThT) fluorescent probe, producing a signal-off readout. The method achieves a detection limit as low as 0.14 fM with excellent single-nucleotide discrimination capability. Its practical utility is validated through recovery experiments, differentiation of cancer cells from normal cells, and high consistency with PCR-based methods in clinical tissue samples. This Cb Ago-Cas9n-EXPAR platform offers a robust and isothermal tool for early diagnosis of TP53-mutated colorectal cancer and can be readily reprogrammed for other clinically relevant single-nucleotide variants. Download: Download high-res image (123KB) Download: Download full-size image
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高影响因子
BIOMATERIALS
·
2020
·
IF 10.317
Chengli Yang,Yuyin Fu,Cheng Huang,Danrong Hu,Kai Zhou,Ying Hao,Bingyang Chu,Yun Yang,Zhiyong Qian
Photodynamic therapy (PDT) is a relatively safe and clinically promising treatment to combat primary tumors, especially epidermal carcinoma, while has negligible effects on distant metastasis. Therefore, this work reports a multifunctional nanosystem ( [email protected] ) exerting a combined photodynamic and immunotherapy to amplify the therapeutic effect on primary tumors and distant metastasis. Specifically, this nanosystem was obtained by electrostatic adsorption of a negatively charged hyaluronic acid “shell” with a positively charged “core” consisting of the CRISPR-Cas9 system targeting the Ptpn2 gene (Cas9-Ptpn2) and a modified mitochondria-targeting chlorin e6 (TPP-PEI-Ce6). Cell experiments demonstrated that the [email protected] nanoparticles possessed very high transfection efficiency on B16F10 cells, and TPP-PEI-Ce6 in the nanoparticles resulted in a significant PDT efficacy due to the efficient singlet oxygen generation in mitochondria under laser-irradiation. The accumulation of the nanoparticles in the tumor by active and passive tumor-targeting in vivo led to the disruption of the Ptpn2 gene by the Cas9-Ptpn2 plasmids in the nanocarriers, thus sensitizing tumors to immunotherapy by the increase of the IFN-γ and TNF-α signaling and the promotion of the proliferation of CD8 + T cells . In addition, Hyaluronidase was administered in advance to destroy the hyaluronic acid in the condensed extracellular matrix and to remove the hyaluronic acid “shell” from the nanosystem, subsequently leading to an enhanced penetration of oxygen and therapeutic agents. Fortunately, the primary and distant tumors in the experimental animals were remarkably inhibited after the combination of PDT-immunotherapy, thus, this easy-to-built nanomedicine could be used as a potential combination therapy against tumors.
研究领域:
癌症治疗
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纳米技术
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分子生物学
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免疫学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2026
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IF 7.8
Yuduo Gao,Yiqing Chen,Zhenghao Hu,Ziyue Wang,Dongchang Sun,Chengfan Wu,Qi Shuai,Yunfeng Yan
Heat shock protein 47 (HSP47) regulates collagen synthesis and fibrosis, which makes it a promising target for controlling pulmonary fibrosis. However, challenges such as cell-selective and efficient drug delivery continue to impede the verification and translation of HSP47-targeted therapies. The CRISPR/Cas9 system can effectively inhibit the overexpression of HSP47 within fibrotic lesions while significantly reducing off-target effects. This study develops a class of lipid nanoparticles (LNPs) capable of codelivering Cas9 mRNA and sgHSP47 to fibroblasts. By incorporation of an additional cationic effector molecule, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), these LNPs achieve differential delivery across various cell lines and markedly enhance uptake efficiency in fibroblasts. This system mediates effective HSP47 knockdown in L929 cells and the lungs of C57BL/6 mice, leading to reduced collagen deposition and fibroblast activation in bleomycin-induced idiopathic pulmonary fibrosis (IPF) mice. This LNP platform holds substantial potential for gene editing-based IPF therapy and provides valuable insights for the efficient and selective delivery of CRISPR/Cas9 systems.
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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
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免疫治疗
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纳米医学
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基因编辑
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癌症生物学
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肿瘤微环境
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DOI
PubMed
高影响因子
AQUACULTURE
·
2025
·
IF 3.9
Tianyang Sun,Xinzhan Meng,Yaru Li,Hongquan Zhang,Ruitao He,Ruicong He,Tiegang Liu,Wenyao Lv,Xiaoyan Xu,Yubang Shen,Jianfeng Ren,Jiale Li
Aquaculture is critical for global food security, yet genetic improvement in farmed fish has progressed more slowly than in terrestrial livestock. Myostatin (MSTN), a conserved negative regulator of skeletal muscle growth in vertebrates, presents a prime target for enhancing aquaculture productivity. In this study, we employed CRISPR/Cas9 ribonucleoprotein (RNP) complexes to disrupt mstnb , the orthologous gene of livestock mstn , in grass carp ( Ctenopharyngodon idella ), the most economically important freshwater aquaculture species in China. Co-injection of five single-guide RNAs (sgRNAs) targeting mstnb exon 1 with Cas9 protein into single-cell embryos achieved 62.0–97.8 % editing efficiency in F 0 mosaic individuals, with an overall population efficiency of 67.1 %. At 7 months post-hatching, mstnb -knockout (KO) fish displayed 17.5 % greater body weight, 8.8 % increased body width, and 7.6 % higher body height compared to control siblings. Histological analysis revealed marked muscle fiber hypertrophy, with cross-sectional area increases of 9.0–26.3 % in cephalosome, trunk, and rump muscles. KO fish also displayed reduced hepatic lipid and glycogen accumulation, indicative of improved metabolic efficiency. While muscle amino acid composition remained largely unchanged, we observed a modest decrease in ω-3 polyunsaturated fatty acids (e.g., EPA and DHA). This study marks the first successful application of CRISPR/Cas9 in grass carp, demonstrating that mstnb disruption enhances growth and muscle development without substantially compromising meat quality. Our results underscore the potential of genome editing for sustainable aquaculture breeding, offering a promising strategy to address rising global protein demands.
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高影响因子
VIROLOGY
·
2023
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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
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2025
·Vol. 56
·期 1
·1-14
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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
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兽医学
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PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
·
2019
·Vol. 47
·期 12
·1425-1432
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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
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分子生物学
·
行为科学
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PubMed
高影响因子
Advanced Science
·
2026
·
IF 14.1
Zi-Jian Huang,Feng-Ming Li,Yi-Fan Tu,Ke-Ke Feng,Cheng-Lei Li,Shi-Cheng Tian,Yong-Shan Hu,Jing-Wei Shao,Zhen-Hua Liu
Photothermal therapy (PTT), a non-invasive tumor treatment, shows promise but is limited in solid tumors by restricted tissue penetration, thermotolerance, anti-apoptotic and immunosuppressive effects. In this study, tumor microenvironment-responsive nanoplatform VARH was constructed based on MXene. Under NIR-II laser irradiation, VARH achieves a high photothermal conversion efficiency of 44.21%. Loaded 2,2'-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride decomposes at high temperatures to generate alkyl radicals, synergizing with hydroxyl radicals from V 4+ -catalyzed endogenous H 2 O 2 decomposition, enabling chemodynamic therapy (CDT) and thermal dynamic therapy to enhance tumor cell oxidative damage. Triggered by high glutathione, VARH releases ribonucleoprotein (RNP) complexes to knockout heat shock protein 90 (HSP90), attenuating cellular heat resistance and promoting apoptosis. It also enhances T cell-mediated anti-tumor immunity and, with free radicals, promotes tumor cell immunogenic cell death (ICD), achieving immunotherapeutic multi-effect synergy. Integrating nanotechnology with precise gene editing, this study develops a novel multimodal synergistic therapy system, providing new insights for multi-modal treatment R&D and advancing PTT and free radical-based cancer therapies.
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PubMed
高影响因子
BIOMATERIALS
·
2025
·
IF 12.9
Ting Fang,Chang Peng,Rui Ding,Yinzhen Fan,Jiajia Jia,Jiajie Chen,XinHang Zhang,Dongkai Wang,Ji Li
Head and neck squamous cell carcinoma (HNSCC) continues to exhibit a poor prognosis, largely due to late diagnosis and the development of cisplatin resistance. Tumor proliferation in HNSCC is closely associated with upregulation of key glycolytic enzymes. However, monotherapeutic targeting of glycolysis paradoxically enhances compensatory glutaminolysis via glutamate overproduction. To overcome this metabolic adaptation, we developed biomimetic nanoparticles P-T-p@CM, fabricated from the pH/reduction dual-responsive copolymer poly(lactic acid)-polyhistidine-polyethylenimine (PLA-pHis-ss-PEI). This nanoplatform enables coordinated co-delivery of telaglenastat (a glutaminase 1 (GLS1) inhibitor) and a CRISPR-Cas9 plasmid encoding sgRNA targeting HIF-1α. This system utilizes homologous cancer cell membrane coating for precise tumor homing, with stimuli-responsive release enabling simultaneous dual metabolic blockade: CRISPR-mediated HIF-1α knockout attenuates glycolysis while telaglenastat suppresses glutamine-to-glutamate conversion. Metabolic analyses confirmed significant reduction in Glycolysis proton efflux rate (GlycoPER), the oxygen consumption rate (OCR) and ATP generation, as well as the related metabolites including the lactate production and glutamate. This dual-starvation strategy depleted energy reserves and biosynthetic precursors, inducing severe metabolic disruption. Notably, in vivo studies showed a 90% tumor inhibition rate (TIR) after 15 days of treatment, through enhanced apoptosis, reduced proliferation, and tumor glucose/glutamate depletion. Collectively, P-T-p@CM establishes a paradigm-shifting approach to disrupt metabolic compensation in the treatment of HNSCC.
研究领域:
肿瘤学
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纳米医学
·
癌症代谢
·
基因治疗
·
药物递送系统
·
分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF CONTROLLED RELEASE
·
2023
·
IF 10.8
Xiujuan Fu,Jianbin Li,Yue Wu,Canquan Mao,Yuhong Jiang
Metastasis is one of the most significant causes for deterioration of breast cancer, contributing to the clinical failure of anti-tumour drugs. Excessive inflammatory responses intensively promote the occurrence and development of tumour, while protease-activated receptor 2 (PAR2) as a cell membrane receptor actively participates in both tumour cell functions and inflammatory responses. However, rare investigations linked PAR2-mediated inflammatory environment to tumour progression. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology is an emerging and powerful gene editing technique and can be applied for probing the new role of PAR2 in breast cancer metastasis, but it still needs the development of an efficient and safe delivery system. This work constructed anionic bovine serum albumin (BSA) nanoparticles to encapsulate CRISPR/Cas9 plasmid encoding PAR2 sgRNA and Cas9 (tBSA/Cas9-PAR2) for triggering PAR2 deficiency. tBSA/Cas9-PAR2 remarkably promoted CRISPR/Cas9 to enter and transfect both inflammatory and cancer cells, initiating precise PAR2 gene editing in vitro and in vivo . PAR2 deficiency by tBSA/Cas9-PAR2 effectively suppressed NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome signalling in inflammatory microenvironment to magnify stimulator of interferon genes (STING) signalling, reactive oxygen species (ROS) accumulation and epithelial-mesenchymal transition (EMT) reversal, consequently preventing breast cancer metastasis. Therefore, this study not only demonstrated the involvement and underlying mechanism of PAR2 in tumour progression via modulating inflammatory microenvironment, but also suggested PAR2 deficiency by tBSA/Cas9-PAR2 as an attractive therapeutic strategy candidate for breast cancer metastasis.
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PubMed
高影响因子
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