Nature Communications
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2025
·Vol. 16
·期 1
·1-19
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IF 15.7
Li Shaoyue,Yu Jifeng,Shen Yuting,Xiong Bing,Zhao De,Xu Weichen,Zhang Shen,Guan Xin,Liu Yunyun,Shan Xuexia,Zhu Anqi,Lyu Qi,Fang Yan,Chen Zitong,Yin Haohao,Sun Liping,Xu Huixiong
Obesity, a surging global health challenge, necessitates effective, accessible and innovative therapeutic models. Here we develop a spatiotemporally controllable microneedle (MN) drug delivery platform for sono-gene therapy to fight obesity. The platform delivers the methoxy polyethylene glycol-polyethyleneimine (mPEG-PEI) modified metal-organic frameworks (MOFs) sonosensitizer and the clustered regularly interspaced short palindromic repeats-activating (CRISPRa)/CRISPRa-uncoupling protein 1 (UCP1) system intradermally to adipocytes. Overall, this therapy platform is capable of achieving two major strategies of “annihilation” and “countermeasure”: one is to kill redundant white adipocytes by sonodynamic therapy, and the other is to promote the browning of white adipocytes through the controllable release of CRISPRa-UCP1 system and sonodynamic effect. Obese male mice treated with this sono-gene therapy shows significant ameliorate in glucose tolerance and insulin sensitivity, successfully achieves weight loss and restrains weight rebound. This study may enable a standard treatment paradigm for sono-gene therapy of obesity and other metabolic diseases.
研究领域:
医学
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生物技术
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药剂学
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PubMed
高影响因子
ACS Applied Materials & Interfaces
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2025
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IF 8.2
Xuekun Fu,Chuanxin Zhong,Wentao Liu,Fuzeng Ren,Chao Liang
Osteoarthritis (OA) is a common joint disorder that causes significant disability. Previous studies suggested that the predominance of M1 macrophages (MΦs) exacerbates inflammation and cartilage degradation in OA, suggesting that shifting the polarization toward M2 MΦs could be a promising therapeutic strategy. We recently developed CRISPRa-engineered macrophages, termed Elite MΦs, that express IL-10 and maintain a stable M2 phenotype. However, achieving effective and sustained delivery of these cells to the OA joint remains a challenge. In this study, we synthesized two injectable aldehyde hyaluronic acid-based hydrogels, CHO/CDH and ACHO/CDH hydrogels, to serve as Elite MΦ delivery platforms. Comprehensive analyses identified the ACHO/CDH hydrogel as superior due to its enhanced suitability for encapsulating and delivering Elite MΦs. When loaded with Elite MΦs, the ACHO/CDH hydrogel was able to not only localize Elite MΦs but also enhance their anti-inflammatory and reparative effects. Furthermore, intra-articular injection of the Elite MΦ-loaded ACHO/CDH hydrogel in an OA mouse model resulted in notable improvements in the joint’s cellular environment, alleviating cartilage degradation and synovial inflammation. These results highlight the ability of the ACHO/CDH hydrogel to rebalance the inflammatory imbalance and promote cartilage repair. This approach not only targets the underlying inflammatory processes more directly than traditional therapies but also harnesses the regenerative potential of macrophages, offering a transformative strategy for OA management.
研究领域:
医学
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生物材料学
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基因编辑技术
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PubMed
高影响因子
BIORESOURCE TECHNOLOGY
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2025
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IF 9
Yang Li,Changyang Yang,Mingxiong Liu,Hongxin Fu,Jufang Wang
Biomanufacturing offers a promising alternative for the production of the building block of nylons, of which butyrolactam is an important platform chemical. To date, there is few studies on butyrolactam biosynthesis, and static control characterized by orientation, irreversibility and ineffectiveness often impedes synthesis efficiency. Here, the chassis Escherichia coli was systematically engineered for butyrolactam synthesis by stepwise optimization strategies. In particular, inadequate α-ketoglutarate pool was identified as the main bottleneck, and then the CRISPRa and CRISPRi served as ON- and OFF-switch for establishing the temperature-powered bioswitch (thereafter renamed AITB circuit), achieving simultaneous transcriptional down- and up-regulation of the multi-genes. In the fed-batch culture, the highest titer of 59.87 g/L butyrolactam was produced by redirecting α-ketoglutarate catabolism. Further, with cassava starch as the substrate, a total of 14.96 g/L butyrolactam was generated, providing the possibility for bioproduct synthesis from renewable feedstock. This study provides efficient tool for developing high-performance biorefineries for bio-based compounds production of interest.
研究领域:
合成生物学
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代谢工程
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生物技术
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DOI
PubMed
高影响因子
Biotechnology Journal
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2025
·Vol. 20
·期 6
·e70036
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IF 3.1
Shupeng Ruan,Aoxue Wang,Hongyi Zou,Ying Lin,Lei Ye,Shuli Liang
The CRISPR activation (CRISPRa) transcriptional system has become a powerful synthetic biology tool for the regulation of endogenous gene expression, allowing for precise fine-tuning of target genes through the simple modification of sgRNA sequences. In this study, we demonstrate that sgRNAs can be effectively expressed using the T7 transcription system. The insertion of tRNA sequences between PT7 and sgRNAs significantly enhances the efficiency of transcriptional activation. Furthermore, the design of PT7-tRNA-sgRNA arrays facilitates the multiplexed activation of genes. sgRNA expression was regulated by the Tet-on induction system, split-T7 system, and RNA cleavage processing by HH-HDV, resulting in the creation of a Boolean logic gene circuit capable of performing both AND and OR logic operations. Finally, we developed a UPR self-responsive system by utilizing endogenous promoters that are responsive to UPR signals to control the expression of T7 RNAP. This system dynamically regulates the expression of the endogenous HAC1 transcription factor, thus enhancing the secretion of heterologous proteins. The findings from this study highlight the potential of utilizing the T7 transcription system for the construction of genetic circuits, providing a practical toolkit for gene regulation in the industrial Komagataella phaffii strain.
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高影响因子
Acta Biomaterialia
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2023
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IF 9.7
Chuanxin Zhong,Shan He,Yuhong Huang,Jianfeng Yan,Junqin Wang,Wentao Liu,Ju Fang,Fuzeng Ren
Clustered regularly interspaced short palindromic repeat activation (CRISPRa) technology has emerged as a precise genome editing tool for activating endogenous transgene expression. While it holds promise for precise cell modification, its translation into tissue engineering has been hampered by biosafety concerns and suboptimal delivery methods. To address these challenges, we have developed a CRISPRa non-viral gene delivery platform by immobilizing non-viral CRISPRa complexes into a biocompatible hydrogel/nanofiber (Gel/NF) composite scaffold. The Gel/NF scaffold facilitates the controlled and sustained release of CRISPRa complexes and also promotes cell recruitment to the scaffold for efficient and localized transfection. As a proof of concept, we employed this CRISPRa delivery platform to activate the vascular endothelial growth factor (VEGF) gene in a rat model with full-thickness skin defects. Our results demonstrate sustained upregulation of VEGF expression even at 21 days post-implantation, resulting in enhanced angiogenesis and improved skin regeneration. These findings underscore the potential of the Gel/NF scaffold-based CRISPRa delivery platform as an efficient and durable strategy for gene activation, offering promising prospects for tissue regeneration. Statement of significance Translation of clustered regularly interspaced short palindromic repeat activation (CRISPRa) therapy to tissue engineering is limited by biosafety concerns and unsatisfactory delivery strategy. To solve this issue, we have developed a CRISPRa non-viral gene delivery platform by immobilizing non-viral CRISPRa complexes into a biocompatible hydrogel/nanofiber (Gel/NF) composite scaffold. This scaffold enables controlled and sustained release of CRISPRa and can induce cell recruitment for localized transfection. As a proof of concept, we activated vascular endothelial growth factor (VEGF) in a rat model with full-thickness skin defects, leading to sustained upregulation of VEGF expression, enhanced angiogenesis and improved skin regeneration in vivo . These findings demonstrate the potential of this platform for gene activation, thereby offering promising prospects for tissue regeneration.
研究领域:
生物技术
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组织工程
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基因治疗
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再生医学
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PubMed
高影响因子
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