Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion
Nuclear protein delivery underlies an array of biotechnological and therapeutic applications. While many variations of protein delivery methods have been described, it can still be difficult or inefficient to introduce exogenous proteins into plants. A major barrier to progress is the cell wall whic...
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MDPI AG
2024-12-01
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| Series: | Current Issues in Molecular Biology |
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| Online Access: | https://www.mdpi.com/1467-3045/46/12/870 |
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| author | Qufei Gu Nathan Ming Yalikunjiang Aizezi Xiaoyang Wei Yizhong Yuan Brian Esquivel Zhi-Yong Wang |
| author_facet | Qufei Gu Nathan Ming Yalikunjiang Aizezi Xiaoyang Wei Yizhong Yuan Brian Esquivel Zhi-Yong Wang |
| author_sort | Qufei Gu |
| collection | DOAJ |
| description | Nuclear protein delivery underlies an array of biotechnological and therapeutic applications. While many variations of protein delivery methods have been described, it can still be difficult or inefficient to introduce exogenous proteins into plants. A major barrier to progress is the cell wall which is primarily composed of polysaccharides and thus only permeable to small molecules. Here, we report a partial enzymatic cell wall digestion-mediated uptake method that efficiently delivers protein into the nucleus of plant cells. Such a method allowed efficient nuclear delivery of green fluorescent protein (GFP) flanked by two nuclear localization sequences (NLS) into <i>Arabidopsis thaliana</i> epidermal root cells without the usual need for large doses of nanoparticles or tissue cultures. We also show that switching from daylight to far-red light-grown conditions promotes effective protein penetration into deep cell layers. This study establishes that a partial enzymatic cell wall degradation could be applied to other cell organelles by changing the localization sequence, paving the way toward the rational engineering of plants. |
| format | Article |
| id | doaj-art-9d7929a253a54960ba3729a458f2219c |
| institution | Kabale University |
| issn | 1467-3037 1467-3045 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Current Issues in Molecular Biology |
| spelling | doaj-art-9d7929a253a54960ba3729a458f2219c2024-12-27T14:18:22ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452024-12-014612144871449610.3390/cimb46120870Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall DigestionQufei Gu0Nathan Ming1Yalikunjiang Aizezi2Xiaoyang Wei3Yizhong Yuan4Brian Esquivel5Zhi-Yong Wang6Department of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USADepartment of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USADepartment of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USADepartment of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USADepartment of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USADepartment of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USADepartment of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305, USANuclear protein delivery underlies an array of biotechnological and therapeutic applications. While many variations of protein delivery methods have been described, it can still be difficult or inefficient to introduce exogenous proteins into plants. A major barrier to progress is the cell wall which is primarily composed of polysaccharides and thus only permeable to small molecules. Here, we report a partial enzymatic cell wall digestion-mediated uptake method that efficiently delivers protein into the nucleus of plant cells. Such a method allowed efficient nuclear delivery of green fluorescent protein (GFP) flanked by two nuclear localization sequences (NLS) into <i>Arabidopsis thaliana</i> epidermal root cells without the usual need for large doses of nanoparticles or tissue cultures. We also show that switching from daylight to far-red light-grown conditions promotes effective protein penetration into deep cell layers. This study establishes that a partial enzymatic cell wall degradation could be applied to other cell organelles by changing the localization sequence, paving the way toward the rational engineering of plants.https://www.mdpi.com/1467-3045/46/12/870green fluorescent proteinnuclear localization sequencepeptidecell wallpartial enzymatic digestion |
| spellingShingle | Qufei Gu Nathan Ming Yalikunjiang Aizezi Xiaoyang Wei Yizhong Yuan Brian Esquivel Zhi-Yong Wang Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion Current Issues in Molecular Biology green fluorescent protein nuclear localization sequence peptide cell wall partial enzymatic digestion |
| title | Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion |
| title_full | Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion |
| title_fullStr | Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion |
| title_full_unstemmed | Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion |
| title_short | Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion |
| title_sort | direct nuclear delivery of proteins on living plant via partial enzymatic cell wall digestion |
| topic | green fluorescent protein nuclear localization sequence peptide cell wall partial enzymatic digestion |
| url | https://www.mdpi.com/1467-3045/46/12/870 |
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