Generation of induced pluripotent stem （iPS） cells from somatic cells has been achieved successfully by simultaneous viral transduction of defined reprogramming transcription factors （TFs）. However, the process re...Generation of induced pluripotent stem （iPS） cells from somatic cells has been achieved successfully by simultaneous viral transduction of defined reprogramming transcription factors （TFs）. However, the process requires multiple viral vectors for gene delivery. As a result, generated iPS cells harbor numerous viral integration sites in their genomes. This can increase the probability of gene mutagenesis and genomic instability, and present significant barriers to both research and clinical application studies of iPS cells. In this paper, we present a simple lentivirus reprogramming system in which defined factors are fused in-frame into a single open reading frame （ORF） via self-cleaving 2A sequences. A GFP marker is placed downstream of the transgene to enable tracking of transgene expression. We demonstrate that this polycistronic expression system efficiently generates iPS cells. The generated iPS cells have normal karyotypes and are similar to mouse embryonic stem cells in morphology and gene expression. Moreover, they can differentiate into cell types of the three embryonic germ layers in both in vitro and in vivo assays. Remarkably, most of these iPS cells only harbor a single copy of viral vector. This system provides a valuable tool for generation of iPS cells, and our data suggest that the balance of expression of transduced reprogramming TFs in each cell is essential for the reprogramming process. More importantly, when delivered by non-integrating gene-delivery systems, this re-engineered single ORF will facilitate efficient generation of human iPS cells free of genetic modifications.展开更多
Development of tools for targeted modifications of specific DNA sequences in plants is of great importance to basic plant biology research as well as crop improvement.The ability to cut DNA at specific locations in th...Development of tools for targeted modifications of specific DNA sequences in plants is of great importance to basic plant biology research as well as crop improvement.The ability to cut DNA at specific locations in the genome to generate doublestrand breaks（DSBs）in vivo is a prerequisite for any genome editing efforts.展开更多
Triple-negative breast cancer(TNBC)is a highly aggressive malignancy predominantly managed via chemotherapy.Our clinical sample analysis revealed a significant correlation between elevated CD24 expression in TNBC tumo...Triple-negative breast cancer(TNBC)is a highly aggressive malignancy predominantly managed via chemotherapy.Our clinical sample analysis revealed a significant correlation between elevated CD24 expression in TNBC tumor cells and patient survival rates.We developed a novel antibody-drug conjugate(ADC),named HN03,consisting of an antibody with engineered cysteines for site-specific conjugation with a low toxic nitric oxide(NO)precursor as its payload through a novel Pt(IV)-mediated bioorthogonal self-cleavable linker.HN03 specifically targets tumor cells expressing high levels of CD24,concurrently generating cisplatin and releasing NO upon activation.HN03 also exhibited potent in vitro and in vivo antitumor activity.It significantly reduced tumor growth at various doses,prevented tumor metastasis,with markedly lower toxicity than traditional chemotherapy agents.We found that a key mechanism of its action involved inducing apoptosis and endoplasmic reticulum stress,substantially decreasing the number of M2-type macrophages.Overall,HN03 stands out as a promising therapeutic option for TNBC,offering a targeted treatment with reduced side effects and the potential for improved outcomes.Furthermore,using Pt(IV)in the linker and an NO precursor as the payload enhances the versatility of the Antibody-NO donor Conjugate(ANC),offering new avenues for the design of the next generation of ADCs.展开更多
文摘Generation of induced pluripotent stem （iPS） cells from somatic cells has been achieved successfully by simultaneous viral transduction of defined reprogramming transcription factors （TFs）. However, the process requires multiple viral vectors for gene delivery. As a result, generated iPS cells harbor numerous viral integration sites in their genomes. This can increase the probability of gene mutagenesis and genomic instability, and present significant barriers to both research and clinical application studies of iPS cells. In this paper, we present a simple lentivirus reprogramming system in which defined factors are fused in-frame into a single open reading frame （ORF） via self-cleaving 2A sequences. A GFP marker is placed downstream of the transgene to enable tracking of transgene expression. We demonstrate that this polycistronic expression system efficiently generates iPS cells. The generated iPS cells have normal karyotypes and are similar to mouse embryonic stem cells in morphology and gene expression. Moreover, they can differentiate into cell types of the three embryonic germ layers in both in vitro and in vivo assays. Remarkably, most of these iPS cells only harbor a single copy of viral vector. This system provides a valuable tool for generation of iPS cells, and our data suggest that the balance of expression of transduced reprogramming TFs in each cell is essential for the reprogramming process. More importantly, when delivered by non-integrating gene-delivery systems, this re-engineered single ORF will facilitate efficient generation of human iPS cells free of genetic modifications.
基金supported by a National Transgenic Science and Technology Program (2016ZX08010002)to R.W.a startup fund from the Huazhong Agricultural University
文摘Development of tools for targeted modifications of specific DNA sequences in plants is of great importance to basic plant biology research as well as crop improvement.The ability to cut DNA at specific locations in the genome to generate doublestrand breaks（DSBs）in vivo is a prerequisite for any genome editing efforts.
基金supported by grants from the National Natural Science Foundation of China(82173681,82104004,81973223 and 82273780)supported by grants from the Natural Science Foundation of Xinjiang Uygur Autonomous Region(2022D01D38 and 2023D01D16,China)+5 种基金open project of the State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia Fund(SKL-HIDCA-2024-4,China)Xinjiang Uygur Autonomous Region Science and Technology Support Project(2022E02123,China)Xinjiang Uygur Autonomous Region Tianshan Talent Training Program(2023TSYCCX0071,China)the Fundamental Research Funds for the Central Universities(No.2632023TD04,China)“Double First-Class”University projects CPU2022PZQ15 from China Pharmaceutical Universitysupported by the National College Students Innovation and Entrepreneurship Training Program(202210316062Y).
文摘Triple-negative breast cancer(TNBC)is a highly aggressive malignancy predominantly managed via chemotherapy.Our clinical sample analysis revealed a significant correlation between elevated CD24 expression in TNBC tumor cells and patient survival rates.We developed a novel antibody-drug conjugate(ADC),named HN03,consisting of an antibody with engineered cysteines for site-specific conjugation with a low toxic nitric oxide(NO)precursor as its payload through a novel Pt(IV)-mediated bioorthogonal self-cleavable linker.HN03 specifically targets tumor cells expressing high levels of CD24,concurrently generating cisplatin and releasing NO upon activation.HN03 also exhibited potent in vitro and in vivo antitumor activity.It significantly reduced tumor growth at various doses,prevented tumor metastasis,with markedly lower toxicity than traditional chemotherapy agents.We found that a key mechanism of its action involved inducing apoptosis and endoplasmic reticulum stress,substantially decreasing the number of M2-type macrophages.Overall,HN03 stands out as a promising therapeutic option for TNBC,offering a targeted treatment with reduced side effects and the potential for improved outcomes.Furthermore,using Pt(IV)in the linker and an NO precursor as the payload enhances the versatility of the Antibody-NO donor Conjugate(ANC),offering new avenues for the design of the next generation of ADCs.
