ACS Central Science
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2023
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IF 18.2
Zhicheng Le,Qi Pan,Zepeng He,Hong Liu,Yi Shi,Lixin Liu,Zhijia Liu,Yuan Ping,Yongming Chen
Intracellular delivery of therapeutic biomacromolecules is often challenged by the poor transmembrane and limited endosomal escape. Here, we establish a combinatorial library composed of 150 molecular weight-defined gemini amphiphiles (GAs) to identify the vehicles that facilitate robust cytosolic delivery of proteins in vitro and in vivo. These GAs display similar skeletal structures but differential amphiphilicity by adjusting the length of alkyl tails, type of ionizable cationic heads, and hydrophobicity or hydrophilicity of a spacer. The top candidate is highly efficient in translocating a broad spectrum of proteins with various molecular weights and isoelectric points into the cytosol. Particularly, we notice that the entry mechanism is predominantly mediated via the lipid raft-dependent membrane fusion, bypassing the classical endocytic pathway that limits the cytosolic delivery efficiency of many presently available carriers. Remarkably, the top GA candidate is capable of delivering hard-to-deliver Cas9 ribonucleoprotein in vivo, disrupting KRAS mutation in the tumor-bearing mice to inhibit tumor growth and extend their survival. Our study reveals a GA-based small-molecule carrier platform for the direct cytosolic delivery of various types of proteins for therapeutic purposes.
研究领域:
药学
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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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药物递送系统
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PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2022
·Vol. 20
·期 1
·1-16
·
IF 9.429
Wang Dan,Chen Ling,Li Chengbi,Long Quanxin,Yang Qing,Huang Ailong,Tang Hua
Background Currently, there are no curative drugs for hepatitis B virus (HBV). Complete elimination of HBV covalently closed circular DNA (cccDNA) is key to the complete cure of hepatitis B virus infection. The CRISPR/Cas9 system can directly destroy HBV cccDNA. However, a CRISPR/Cas9 delivery system with low immunogenicity and high efficiency has not yet been established. Moreover, effective implementation of precise remote spatiotemporal operations in CRISPR/Cas9 is a major limitation.Results In this work, we designed NIR-responsive biomimetic nanoparticles (UCNPs-Cas9@CM), which could effectively deliver Cas9 RNP to achieve effective genome editing for HBV therapy. HBsAg, HBeAg, HBV pgRNA and HBV DNA along with cccDNA in HBV-infected cells were found to be inhibited. These findings were confirmed in HBV-Tg mice, which did not exhibit significant cytotoxicity and minimal off-target DNA damage.Conclusion sThe UCNPs-based biomimetic nanoplatforms achieved the inhibition of HBV replication via CRISPR therapy and it is a potential system for efficient treatment of human HBV diseases.Graphical Abstract
研究领域:
分子生物学
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病毒学
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纳米技术
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治疗学
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PubMed
高影响因子
ACS Applied Materials & Interfaces
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2024
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IF 8.3
Lingjiao Li,Chengyao Feng,Wenchao Zhang,Lin Qi,Binfeng Liu,Hua Wang,Chenbei Li,Zhihong Li,Chao Tu,Wenhu Zhou
Cisplatin, a frontline chemotherapeutic agent against cancer, faces challenges in clinical application due to significant toxicities and suboptimal efficacy. Renal toxicity, a dose-limiting factor of cisplatin, results from multifactorial processes including cisplatin-induced cellular pyroptosis, oxidative damage, and inflammatory responses. Our findings reveal that Tea Polyphenols Nanoparticles (TPNs) derived from Epigallocatechin gallate (EGCG) effectively could address these diverse mechanisms, comprehensively alleviating cisplatin-induced nephrotoxicity. Leveraging TPNs as carriers, chemical conjugation enables the encapsulation of tetravalent cisplatin prodrug, extending its systemic half-life, enhancing tumor tissue accumulation, while simultaneously mitigating renal toxicity. Concurrently, employing a CRISPR/Cas9 kinase library, we identified CSNK2A1 as a target sensitizing tumor cells to cisplatin, enabling specific siRNA sequences to augment cisplatin susceptibility, thereby minimizing the dosage requirement. Benefiting from the versatile carrier properties of TPNs to codeliver cisplatin prodrug and anti-CSNK2A1 siRNA, we developed a codelivery system, Pt-TPNs/siRNA. Pt-TPNs/siRNA not only enhances the anticancer effects but also mitigates cisplatin-induced renal toxicity, achieving efficacy while reducing toxicity. Mechanistic and safety assessments of these nanoparticles were conducted at both cellular and animal levels, opening new avenues for improved clinical utilization of cisplatin.
研究领域:
肿瘤学
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药理学
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纳米技术
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分子生物学
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毒理学
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DOI
PubMed
高影响因子
Nature Communications
·
2022
·Vol. 13
·期 1
·1-16
·
IF 17.694
Yang Bin,Kong Jilie,Fang Xueen
Recent advances in biointerfaces have led to the development of wearable devices that can provide insights into personal health. As wearable modules, microneedles can extract analytes of interest from interstitial fluid in a minimally invasive fashion. However, some microneedles are limited by their ability to perform highly effective extraction and real-time monitoring for macromolecule biomarkers simultaneously. Here we show the synergetic effect of CRISPR-activated graphene biointerfaces, and report an on-line wearable microneedle patch for extraction and in vivo long-term monitoring of universal cell-free DNA. In this study, this wearable system enables real-time monitoring of Epstein-Barr virus, sepsis, and kidney transplantation cell-free DNA, with anti-interference ability of 60% fetal bovine serum, and has satisfactory stable sensitivity for 10 days in vivo. The experimental results of immunodeficient mouse models shows the feasibility and practicability of this proposed method. This wearable patch holds great promise for long-term in vivo monitoring of cell-free DNA and could potentially be used for early disease screening and prognosis.
研究领域:
生物技术
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医疗器械
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分子生物学
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可穿戴技术
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基因组学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
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2020
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IF 8.758
Kun Tu,Huan Deng,Li Kong,Yi Wang,Ting Yang,Qian Hu,Mei Hu,Conglian Yang,Zhiping Zhang
Blocking immune checkpoints with monoclonal antibody has been verified to achieve potential clinical successes for cancer immunotherapy. However, its application has been impeded by the “cold” tumor microenvironment. Here, weak acidity-responsive nanoparticles co-loaded with CRISPR/Cas9 and paclitaxel (PTX) with the ability to convert “cold” tumor into “hot” tumor are reported. The nanoparticles exhibited high cargo packaging capacity, superior transfection efficiency, well biocompatibility, and effective tumor accumulation. The CRISPR/Cas9 encapsulated in nanoparticles could specifically knock out cyclin-dependent kinase 5 gene to significantly attenuate the expression of programmed death-ligand 1 on tumor cells. More importantly, PTX co-delivered in nanoparticles could significantly induce immunogenic cell death, reduce regulatory T lymphocytes, repolarize tumor-associated macrophages, and enhance antitumor immunity. Therefore, the nanoparticles could effectively convert cold tumor into hot tumor, achieve effective tumor growth inhibition, and prolong overall survival from 16 to 36 days. This research provided a referable strategy for the development of combinatorial immunotherapy and chemotherapy.
研究领域:
癌症免疫治疗
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纳米技术
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基因编辑
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肿瘤学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
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IF 6.057
Xianfeng Wang,Xiaolong Chen,Chengxiang Chu,Yuanyi Deng,Mei Yang,Danqun Huo,Faliang Xu,Changjun Hou,Jiayi Lv
Accurate and effective detection of single-stranded nucleic acids is vital in both disease diagnosis and pathological studies. Hence, we develop a PAMmer-assisted CRISPR/Cas9 system mediated G4-EXPAR (Cas-G4EX) strategy for site-specific detection of ssRNA and ssDNA . PAMmer-assisted CRISPR/Cas9 executes the site-specific cleavage of target ssRNA or ssDNA and released product fragment with the desired sequence at the 3′-terminal. This fragment serves as a primer to activate subsequent sequence-dependent exponential amplification reaction (EXPAR). The G-rich EXPAR products assembles with hemin to form a G-Quadruplex (G4/hemin). G4/hemin catalyzes ABTS-H 2 O 2 system with the appearance of vivid green color, realizing naked-eye analysis. Cas-G4EX integrates the superiority of CRISPR/Cas9 and EXPAR, presenting outstanding site-specific recognition and high-performance amplification efficiency. Meanwhile, the programmability of CRISPR/Cas9 system makes the proposed method become a universal detection paradigm for any ssRNA or ssDNA. Cas-G4EX assay shows the linear relationship from 250 aM to 2.5 nM for ssRNA detection with the actual LOD of 250 aM, and that ranges from 100 aM to 1 nM for ssDNA detection with the actual LOD of 100 aM. Additionally, the acceptable recoveries of 101.48%–109.61% for ssRNA and 93.25%–111.98% for ssDNA in real detection of human serum are obtained for detection of single-strand nucleic acid in real samples. Cas-G4EX also exhibits the excellent discrimination for single-base mutation of single-stranded nucleic acids. Therefore, Cas-G4EX assay provides a promising platform in the applications of molecular diagnosis and pathological analysis.
研究领域:
生物技术
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分子生物学
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诊断医学
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病理学
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产品关联
DOI
PubMed
高影响因子
PEST MANAGEMENT SCIENCE
·
2024
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IF 3.8
Yun Wang,Hui-Qian Xu,Hong-Liang Han,Dong Chen,Hongbo Jiang,Guy Smagghe,Jin-Jun Wang,Dong Wei
BACKGROUND The melon fly, Zeugodacus cucurbitae (Coquillett), is an invasive Tephritidae pest with robust fertility. The male accessory glands (MAGs) form a vital organ that ensures insect reproductive efficiency. Most of the secreted proteins by MAGs exhibit a male bias expression. Takeout, one of these proteins, is abundantly present in the MAGs of many insects. RESULTS In this study, we identified 32 takeout genes in Z. cucurbitae . The phylogenetic analysis and multiple sequence alignment results showed that Zctakeout1 is the most related homolog to the MAGs-specific takeout in Tephritidae. The real-time quantitative PCR results showed that Zctakeout1 was exclusively expressed in the male adult stage, and its expression level gradually increased with the increase in age and then remained stable at the sexually matured stage. The distribution among tissues demonstrated the specific expression of Zctakeout1 in the MAGs, and fluorescence immunohistochemical results confirmed the presence of Zctakeout1 in close proximity to binuclear cells of the mesoderm epidermal MAGs. In continuation, CRISPR/Cas9-mediated genome editing was employed, resulting in successfully generating a homozygous strain with an +8 bp insertion. The mating experiments with the Zctakeout1 −/− males resulted in significant reductions in both the mating rate and egg production of females. CONCLUSION These findings prove that the MAGs-specific Zctakeout1 is essential in regulating fecundity in female Z. cucurbitae fruit flies. Our data suggests its utilization in future essential insect-specific gene-directed sterility insect technique (SIT) by the genetic manipulation to keep these important Tephritidae populations under control. © 2024 Society of Chemical Industry.
研究领域:
昆虫学
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遗传学
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分子生物学
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生物技术
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产品关联
DOI
PubMed
高影响因子
Forests
·
2024
·Vol. 15
·期 7
·1157
·
IF 2.4
Huihui Liu,Lin Chen,Qinghua Ma,Xinghong Wang,Sufang Zhang,Xuguo Zhou
Hox genes, specifically the bithorax complex (ubx,abdominal-a, andabdominal-b), play a crucial role in specifying posterior abdominal development and serve as key regulators of germline gene development in insects. However, the function of the bithorax complex in the pine caterpillar moth,Dendrolimus punctatus, a major pine tree defoliator in China, remains largely unknown. Specifically,Abdominal-A(Abd-a) controls regional variation in abdominal segmentation in model insects such asDrosophilaandTribolium; however, its role inD. Punctatusremains unexplored. In this study, CRISPR/Cas9 was used to functionally characterizeAbd-ainD. punctatus. Two target sites were selected, and the genotypes and phenotypes of the G0 and G1 generations were evaluated. Our findings indicate that knocking outAbd-aled to an abnormality in the posterior segments A2–A7, as well as the loss of appendages, mainly prolegs, and affected the thoracic T3 segmentation as well as wing development. Moreover, mutation inAbd-aalso impacted anal and reproductive development. Taken together, these results demonstrate thatDpAbd-ais essential for embryonic and reproductive development inD. punctatusand could be a promising target for genetic control of this devastating conifer defoliator.
研究领域:
昆虫学
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遗传学
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分子生物学
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产品关联
DOI
高影响因子
Aquaculture and Fisheries
·
2026
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IF 5
Weizhong Li,Wenyao Lv,Tianyang Sun,Qian Cong,Weipeng Lian,Tao Sheng,Hongquan Zhang,Qiaozhen Yu,Yuxuan Chen,Xinzhan Meng,Jianfeng Ren,Lang Gui,Junqiang Qiu,Guiling Wang,Xiaoyan Xu,Jiale Li
The transforming growth factor-beta (TGF-beta) signaling pathway is a central regulator of skeletal muscle growth in vertebrates, but the function of Acvr1b (activin receptor type 1B) remains poorly characterized in teleosts. Here, we used CRISPR/Cas9 to disrupt Acvr1b in grass carp ( Ctenopharyngodon idella ) and evaluated growth performance, muscle histology, and myog enic gene expression. Cas9 protein and sgRNAs targeting exons 2 and 3 were co-injected into fertilized eggs, and edited juveniles were identified by T7E1 assay and Sanger sequencing. Acvr1b -edited fish showed significantly improved growth relative to wild-type controls, including a 25.8% increase in body weight and a 7.39% increase in body length. qRT-PCR revealed reduced Acvr1b expression in all examined tissues. Consistent with enhanced muscle growth, mstn was downregulated, myog was upregulated, and myod was unchanged in edited fish. Histological analysis showed increased muscle fiber size and density, indicating that both hypertrophy and hyperplasia contributed to the growth advantage. Together, these findings identify Acvr1b as a promising target for genetic improvement in grass carp aquaculture.
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DOI
高影响因子
Nature Communications
·
2018
·Vol. 9
·期 1
·1-15
·
IF 12.353
Usman Waqas Muhammad,Pham Tin Chanh,Kwok Yuk Yan,Vu Luyen Tien,Ma Victor,Peng Boya,Chan Yuen San,Wei Likun,Chin Siew Mei,Azad Ajijur,He Alex Bai-Liang,Leung Anskar Y. H.,Yang Mengsu,Shyh-Chang Ng,Cho William C.,Shi Jiahai,Le Minh T. N.
Most of the current methods for programmable RNA drug therapies are unsuitable for the clinic due to low uptake efficiency and high cytotoxicity. Extracellular vesicles (EVs) could solve these problems because they represent a natural mode of intercellular communication. However, current cellular sources for EV production are limited in availability and safety in terms of horizontal gene transfer. One potentially ideal source could be human red blood cells (RBCs). Group O-RBCs can be used as universal donors for large-scale EV production since they are readily available in blood banks and they are devoid of DNA. Here, we describe and validate a new strategy to generate large-scale amounts of RBC-derived EVs for the delivery of RNA drugs, including antisense oligonucleotides, Cas9 mRNA, and guide RNAs. RNA drug delivery with RBCEVs shows highly robust microRNA inhibition and CRISPR–Cas9 genome editing in both human cells and xenograft mouse models, with no observable cytotoxicity.
研究领域:
药理学
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分子生物学
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基因治疗
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Wei Chao-Gang,Zhang Rui,Wei Lan-Yi,Pan Peng,Zu He,Liu Ya-Zhen,Wang Yong,Shen Jun-Kang
Introduction: Erythropoietin producing hepatocyte receptor A2 (EphA2) is widely presented in the tumor cells, closely related to tumor cell migration, not cell apoptosis and proliferation. Based on its high expression in castration-resistant prostate cancer (CRPC), we herein develop a CRISPR-Cas9-based genome-editing nanomedicine to target erythropoietin producing hepatocyte receptor A2 for the treatment of castration-resistant prostate cancer.Methods: To this end, TAT was designed to stabilize the distribution of calcium, and then bound to ribonucleoprotein (RNP) to form nanoparticles RNP@CaP-TAT.Results: This nanoparticle has a simple synthesis process with good biocompatible, to achieve the knockout of tumor cells (PC-3) targeting erythropoietin producing hepatocyte receptor A2 gene and to effectively suppress the migration of tumor cells.Discussion: This delivery genome editing system provides a promising gene therapy strategy for the treatment of castration-resistant prostate cancer, showing good potential against castration-resistant prostate cancer tumor metastasis. In addition, it can be extended to other types of cancer with highly heterogeneous gene expression.
研究领域:
纳米技术
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癌症生物学
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基因编辑
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分子生物学
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生物医学工程
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DOI
PubMed
高影响因子
Antibiotics-Basel
·
2026
·Vol. 15
·期 2
·202
·
IF 5.5
Zhiping Hou,Mengliu Peng,Xiaozhi Ju,Yaqi Sun,Liping Du,Ye Liang,Shu-Shan Gao
Background: The fungusAcremonium chrysogenumis crucial for producing cephalosporin antibiotics. While CRISPR/Cas9 has been developed for this species, it has not been applied to first-line industrial strains, to the best of our knowledge. For example, engineering industrialA. chrysogenumfor overproducing deacetoxycephalosporin C (DAOC, an important precursor for clinically used cephalosporin antibiotics) is currently often a multi-step and inefficient process. Methods: Here, we applied CRISPR/Cas9 to create a DAOC overproducer in a single step. Our method uses a donor template to simultaneously delete and overexpress genes, offering a simple, efficient, and time-saving solution. Results: Furthermore, through methodological optimization, the final homozygous multigene-edited strain achieved an industrial-scale DAOC titer of 12.4 ± 0.2 g/L, representing a significant improvement over the initial edited strain (7.2 ± 0.22 g/L). Conclusions: This demonstrates that CRISPR/Cas9 can effectively edit industrialA. chrysogenum, providing a strategy to enhance the production of other cephalosporin precursors.
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高影响因子
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