Nano Today
·
2021
·
IF 20.722
Meng Lin,Zhenzhen Yang,Yiliang Yang,Yiwei Peng,Jiajia Li,Yitian Du,Qi Sun,Datong Gao,Quan Yuan,Yu Zhou,Xinyi Chen,Xianrong Qi
Tumor-associated macrophages (TAMs) are abundant in the tumor microenvironment and promote the tumor progression via multiple mechanisms. CD47 is overexpressed in most malignant tumors and acts as a “don’t eat me” signal to inhibit phagocytosis . We utilized CRISPR/Cas9 technology to knock out CD47 to guarantee the long-lasting anti-tumor immune responses. However, the reprogrammed TAMs are vulnerable to the inhibitory cytokines and tend to be transformed back into TAMs, and CD47 blockade alone may not be sufficient to elicit effective immune responses. We combined CD47 blockade with an immune-activating cytokine IL-12 for synergistic anti-tumor efficacy by genetically engineering the tumor cells into factories of IL-12 to in situ reprogram TAMs. Firstly, we designed a selective responsiveness accelerated gene delivery system named HPT-PFs that was dual modified with hyaluronic acid (HA) and tumor microenvironment sensitive peptides (TMSP) to simultaneously deliver plasmids for CD47 knockout and IL-12 production. Due to tumor-specific transfection and excellent endosome escape ability of HPT-PFs vector, more than 27% of tumor cells lost CD47 expression after HPT-PFs mediated gene editing, thus eliciting the phagocytosis by macrophages. And higher than 500 ng/ml of IL-12 was produced by tumor cells after HPT-PFs mediated pIL-12 expression, indicating the successful engineering of tumor cells into factories of IL-12. In melanoma-bearing mice models, drastic elevation of M1-polarized TAMs and secretion of inflammatory cytokines were observed when combined CD47 knockout with IL-12 production, which led to significant inhibition of tumor growth. Our study suggested that combination of CRISPR-mediated CD47 blockade with IL-12 production in tumor cells could synergistically promote macrophage-mediated immunotherapy, paving the way for CRISPR-based in situ engineering tumor cells for effective immunotherapy.
研究领域:
癌症生物学
·
免疫学
·
基因治疗
·
生物技术
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Kai Shi,Jiaxuan Chen,Ying Cheng,Jiuhua Song,Yang Li,Xin Cheng,Xiaofeng Bai,Jiali Chang,Tao Jiang
Microplastic (MP) pollution has attracted worldwide attention due to its harmful effects on human health and ecosystems. Herein, we develop a novel label-free electrochemical biosensor for sensitive and selective detection of MP based on split gRNA with CRISPR/Cas12a (SCas12a)-mediated cascade strand displacement and an aptamer (termed as CRISPR-MP). Within the CRISPR-MP system, the target polyvinyl chloride (PVC)/polystyrene (PS) triggers the SCas12a-mediated cascade strand displacement, and then activates the DNase activity of SCas12a. Upon cleaving the DNA probe of the G-quadruplex/hemin complex on the gold electrode, the active Cas12a triggers a decrease in the electrochemical signal.This electrochemical aptasensor shows a detection limit of 37 ng/mL of PVC and 45 ng/mL of PS, respectively. The developed CRISPR-MP system expands the utility of the CRISPR for MP detection and exhibits significant promise in protecting human health and monitoring the environment.
研究领域:
生物分析化学
·
环境监测
·
分子诊断学
·
生物传感器技术
·
纳米毒理学
✓ 已验证
产品关联
DOI
高影响因子
ACS Sensors
·
2025
·
IF 9.1
Mengqi Huang,Kunte Shang,Luyi Ying,Yuqi Han,Ningxi Hong,Yu-Feng Yao
Rapid and precise detection of specific DNA is valuable for biological research and clinical disease diagnosis. Clustered regularly interspaced short palindromic repeat (CRISPR) technology can enhance existing DNA testing, making DNA detection faster, more portable, and more accurate. This study presents a new Cas12-driven exponential amplification-based lateral flow biosensor (CADEX-LF) for rapid and highly sensitive DNA detection. CADEX-LF takes full advantage of the highly specific target-loading-dependent trans-cleavage activity of Cas12 and the extremely high efficiency of nicking endonuclease-mediated exponential amplification. The adoption of lateral flow readout enables CADEX-LF for point-of-care (POC) use without requiring complicated supplementary equipment. CADEX-LF was shown to achieve a detection sensitivity of 2 × 10–15 M within 45 min of measurement time and displayed outstanding specificity with double-base resolution. Furthermore, CADEX-LF could identify herpes simplex virus 1 (HSV-1) DNA in tears of rabbits and clinical patients with HSV-1 keratitis, exhibiting its practical application potential in clinical diagnosis. The proposed CADEX-LF biosensor may have great promise for point-of-care disease diagnosis in resource-limited environments.
研究领域:
分子生物学
·
临床诊断学
·
生物传感器技术
·
病毒学
·
精准医学
·
转化医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Synthetic Biology
·
2025
·
IF 3.9
Hanqing Zhang,Luai R. Khoury,Peng Xu
Yarrowia lipolytica, a nonconventional yeast, has become an industrial workhorse to synthesize valuable compounds, including lipids, oleochemicals, and nutraceuticals. While the synthetic biology toolkits to engineer the genome and endogenous metabolic pathways are not as developed as Baker’s yeast, it has emerged as a promising microbial host for industrial applications. In this study, we examined the multiplexed editing capability of the CRISPR-AsCpf1 coupled with gRNAs generated from either a yeast native promoter or an orthogonal T7 promoter, which yielded 73.3% editing efficiency for up to four target genes and 100% editing efficiency for two genes. We also attempted two strategies to enhance homology-directed recombination (HDR) efficiency; only minor improvements were observed. We further demonstrated that CRISPR-dAsCpf1 with T7-driven gRNA achieved significant gene knockdown compared to the CRISPR-RfxCas13d system. Its knockdown efficiency was comparable to that of an antisense T7 promoter system. Taken together, this work provides a facile toolkit that enables efficient and multiplexed genome editing and transcriptional knockdown of critical genes by combining CRISPR-Cpf1 with an orthogonal T7 transcription system in Y. lipolytica.
研究领域:
合成生物学
·
基因工程
·
微生物生物技术
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Jingwen Zhang,Xiru Zhang,Xinna Xie,Biru Han,Feng Zhao,Xinyan Yang,Wei Zhang,Yujun Jiang,Xianlong Zhang
The problem of mycotoxin contamination in foodstuffs has attracted widespread attention and posed a great threat to human health. Therefore, the sensitive and effective detection of mycotoxins is of great importance for preserving public health worldwide. In this study, a CRISPR/Cas12a-mediated marker-free fluorescent biosensing platform was constructed for highly sensitive and fast detection of aflatoxin B1. The copper nanoclusters were synthesized with marker-free DNA single strands within 5 min, showing the outstanding fluorescence properties. With the existence of aflatoxin B1, the released complementary DNA (cDNA) triggered multiple isothermal amplification reaction. Subsequently, the obtained amplification products triggered the trans -cleavage activity of CRISPR/Cas12a system, which degraded the DNA single strands for synthesis of copper nanoclusters, leading to a decreased fluorescent signal. Benefiting from good fluorescence properties of copper nanoclusters, the established biosensing platform for aflatoxin B1 detection had a high specificity and a limit of detection of 47.51 pg/mL in the linear range of 0.05–10 ng/mL. The proposed platform provided a new insight for the detection of non-nucleic acid targets.
✓ 已验证
DOI
PubMed
高影响因子
Advanced Science
·
2022
·Vol. 9
·期 14
·2105231
·
IF 17.521
Fei Zheng,Zhi Chen,Jingfeng Li,Rui Wu,Bin Zhang,Guohui Nie,Zhongjian Xie,Han Zhang
The clustered regularly interspaced short palindromic repeats (CRISPR) molecular system has emerged as a promising technology for the detection of nucleic acids. Herein, the development of a surface plasmon resonance (SPR) sensor that is functionalized with a layer of locally grown graphdiyne film, achieving excellent sensing performance when coupled with catalytically deactivated CRISPR-associated protein 9 (dCas9), is reported. dCas9 protein is immobilized on the sensor surface and complexed with a specific single-guide RNA, enabling the amplification-free detection of target sequences within genomic DNA. The sensor, termed CRISPR-SPR-Chip, is used to successfully analyze recombinant plasmids with only three-base mutations with a limit of detection as low as 1.3 fM. Real-time monitoring CRISPR-SPR-Chip is used to analyze clinical samples of patients with Duchenne muscular dystrophy with two exon deletions, which are detected without any pre-amplification step, yielding significantly positive results within 5 min. The ability of this novel CRISPR-empowered SPR (CRISPR-eSPR) sensing platform to rapidly, precisely, sensitively, and specifically detect a target gene sequence provides a new on-chip optic approach for clinical gene analysis.
研究领域:
分子生物学
·
医学诊断
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Jiaying Wang,Yu Tian,Kai Zhang,Jiantong Jiao,Yi Zhao
NF-κB p50 is a critical transcription factor that plays an essential role in regulating immune responses, inflammatory pathways, and tumor progression. Its aberrant expression is closely linked to various pathological conditions, making it a valuable biomarker for early disease diagnosis and therapeutic monitoring. In this work, we developed a highly sensitive and reusable electrochemical biosensor capable of detecting NF-κB p50 at ultralow concentrations. The biosensor integrates a pH-responsive regeneration mechanism and a multi-stage signal amplification cascade involving Exonuclease III (Exo III)-assisted recycling, CRISPR/Cas12a-mediated trans -cleavage, and DNA tetrahedron nanostructure-functionalized electrodes. This synergistic design significantly enhances detection sensitivity and stability. The platform offers a broad dynamic detection range from 5 aM to 100 fM with a remarkably low detection limit of 2.38 aM. More importantly, the biosensor demonstrates excellent reusability through efficient regeneration under alkaline conditions (pH 10.0), with minimal loss in signal output. Validation in tenfold-diluted human serum samples yielded high recovery rates (99.61 %–106.23 %) and low relative standard deviations, confirming its practical applicability. This study not only presents a robust approach for ultrasensitive protein detection but also introduces a generalizable strategy for the regenerative electrochemical analysis of protein-based biological macromolecules in clinical diagnostics.
研究领域:
生物医学工程
·
分子诊断学
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Jun Wang,Xueting Cui,Jing Zhang,Juan Li,Bo Pang,Jinhua Li
Microcystin-LR (MC-LR) is a significant environmental pollutant that presents a substantial risk to public health. The monitoring with simple approaches is essential for the prevention and management of MC-LR. In this study, we present an aptasensor platform utilizing recombinase polymerase amplification (RPA)-CRISPR/Cas12a technology to accurately identify MC-LR. The MC-LR aptamers were attached to magnetic beads (MBs) following combination with cDNA by hybridization, resulting in the formation of the MB aptasensor. Subsequently, glucose oxidase (GOx)-modified MB were synthesized using single-stranded DNA (MB-ssDNA-GOx) in order to induce alterations in color upon exposure to the TMB-glucose system. The colorimetric signal was visible to the naked eye and exhibited a positive correlation with MC-LR concentration. The sensing platform described in this study demonstrated great sensitivity and achieved a limit of detection of 0.1 μg/L. Furthermore, this methodology exhibited exceptional selectivity and favorable recovery rates, hence showcasing its strong suitability for the analysis of actual water samples. Hence, the assay exhibites significant potential for future environmental monitoring.
研究领域:
环境科学
·
分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
ANALYST
·
2023
·Vol. 148
·期 10
·2308-2315
·
IF 4.2
Ming Zhou,Xin Li,Herui Wen,Bin Huang,Jiali Ren,Jialin Zhang
The simple and efficient detection of nucleic acids is important in the diagnosis of tuberculosis (TB) caused by Mycobacterium tuberculosis (M. tuberculosis). However, base mismatch will lead to false positive and false negative nucleic acid test, which seriously interferes with the accuracy of the final results. Herein, we demonstrated a CRISPR/Cas-9-mediated fluorescent strategy utilizing fluorescence resonance energy transfer (FRET) for the detection of bacteria. High-variable region of M. tuberculosis 16S rDNA fragment was used as the target, and CRISPR/Cas9 was used as the recognition element. The binding of the P1 probe of upconversion nanoparticles (UCNPs) @SiO2-P1 and the P2 probe of Fe3O4@Au-P2 caused the fluorescence quenching of UCNPs. In the presence of the target, the P2 probe hybridized with the target to form double-stranded DNA (dsDNA), which was recognized and cleaved by CRISPR/Cas9, resulting in the breaking of the P1–P2 duplex linkage. UCNPs moved away from Fe3O4@Au under a magnetic field, and the fluorescence signal was restored; bacteria were detected under the excitation of a 980 nm laser source. Using the CRISPR/Cas-9-mediated system, the sensor could distinguish single-base mismatches in 10 bases from the protospacer adjacent motif (PAM) region. The limit of detection (LOD) was 20 CFU mL−1 and the detection time was 2 h. It developed a new way of accurate nucleic acid detection for disease diagnosis.
研究领域:
生物技术
·
医学诊断
·
纳米技术
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2022
·
IF 12.545
Yang Sun,Cheng Yang,Xiaolin Jiang,Pengbo Zhang,Shuo Chen,Fengxia Su,Hui Wang,Weiliang Liu,Xiaofei He,Lei Chen,Baoyuan Man,Zhengping Li
False detection of SARS-CoV-2 is detrimental to epidemic prevention and control. The scalar nature of the detected signal and the imperfect target recognition property of developed methods are the root causes of generating false signals. Here, we reported a collaborative system of CRISPR-Cas13a coupling with the stabilized graphene field-effect transistor, providing high-intensity vector signals for detecting SARS-CoV-2. In this collaborative system, SARS-CoV-2 RNA generates a “big subtraction” signal with a right-shifted feature, whereas any untargets cause the left-shifted characteristic signal. Thus, the false detection of SARS-CoV-2 is eliminated. High sensitivity with 0.15 copies/μL was obtained. In addition, the wide concerned instability of the graphene field-effect transistor for biosensing in solution environment was solved by the hydrophobic treatment to its substrate, which should be a milestone in advancing it's engineering application. This collaborative system characterized by the high-intensity vector signal and amazing stability significantly advances the accurate SARS-CoV-2 detection from the aspect of signal nature.
研究领域:
生物传感
·
纳米技术
·
分子生物学
·
电子学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Baoqing Zhou,Qinghua Ye,Fan Li,Xinran Xiang,Yuting Shang,Chufang Wang,Yanna Shao,Liang Xue,Jumei Zhang,Juan Wang,Yu Ding,Moutong Chen,Qingping Wu
Rapid, accurate point-of-care testing (POCT) for pathogenic bacteria detection is the key for avoiding foodborne diseases caused by pathogens or their toxins. The lateral flow biosensor (LFB) based on clustered regularly interspaced short palindromic repeats (CRISPR/Cas) has displayed remarkable potential for pathogen diagnosis. In this study, we report a novel CRISPR/Cas12a-based fluorescence enhanced LFB in conjunction with functionalized quantum dots , combined with recombinase-assisted amplification (RAA), to establish low-cost, simple, and sensitive detection of Staphylococcus aureus , namely CRA-LFB (CRISPR/Cas-recombinase-assisted amplification based LFB). The CRA-LFB assay is characterized by Cas12a-mediated trans-cleavage activation induced by the target DNA to digest biotin-DNA probes, caused no complementarity to the capture probe immobilized on the test (T) line, and resulted in an undetectable T line fluorescence signal on LFB. The naked eye or fluorescence strip reader was used to determine the fluorescence intensity of the T and control lines. The limit of detection (at optimal conditions) was as low as 75 aM of genomic DNA, and 5.4 × 10 2 cfu/mL of S. aureus in pure cultures were detected. Moreover, this CRA-LFB assay can rapidly and accurately detect S. aureus in spiked and natural meat and vegetable samples. The CRA-LFB method yielded high specificity and no interference from other nontargeted bacteria. The results exhibited high-resolution, high-intensity fluorescent signals obtained within 70 min. This bacterial detection platform is simple, low cost, almost equipment-free, and fully qualified for food and clinical diagnosis onsite testing requirements.
研究领域:
生物技术
·
微生物学
·
食品安全
·
即时诊断
✓ 已验证
DOI
高影响因子
Nano Today
·
2024
·
IF 13.2
Xiuhua Pan,Jie Yang,Ruixin Kang,Jun Zhang,Zongwei Hu,Ning Yang,Zengyi Liu,Zengguang Jia,Xiawei Zhou,Qi Shen
The induction of ferroptosis in cancer cells has emerged as a promising approach to initiate immunogenic cell death (ICD). However, the limited production of lipid peroxidation (LPO) considerably undermines the effectiveness of ferroptosis. In this work, T7 polypeptide engineered bacterial outer membrane vesicles (T7-OMVs) co-delivering plasmids and melanin nanoparticles (MNPs) are fabricated into nanoclusters (TO@Mp) for LPO amplification. The clustered regularly interspaced short palindromic repeats (CRISPR) system as the gene editing tool can knockdown glutathione peroxides 4 ( GPX4 ) to amplify LPO accumulation. Photothermal MNPs exhibit an intrinsic capacity of up-regulating Acyl-CoA synthetase long chain family member 4 (ACSL4) and generating reactive oxygen species (ROS), thereby cooperating with GPX4 gene knockdown to reinforce ferroptosis. While the TO@Mp NPs augment ICD via synergistic lipid peroxidation induced ferroptosis, the mechanism of proinflammatory response acquired by the RNA-seq analysis also indicates that the nanoclusters activate the toll like receptor (TLR) pathways, thereby coordinately potentiating DCs maturation and T cells immunity to restrain tumor growth and metastasis. Conclusively, the biomimetic bacteria-derived platform for synergistic CRISPR-photothermo modulation represents an appealing way for tumor suppression and immunological therapy, which holds great potential in clinical translation for cancer treatment.
研究领域:
癌症生物学
·
免疫治疗
·
纳米技术
·
基因编辑
·
光热疗法
·
生物医学工程
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）