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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Main Authors: Qufei Gu, Nathan Ming, Yalikunjiang Aizezi, Xiaoyang Wei, Yizhong Yuan, Brian Esquivel, Zhi-Yong Wang
Format: Article
Language:English
Published: MDPI AG 2024-12-01
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
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institution Kabale University
issn 1467-3037
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publishDate 2024-12-01
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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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