In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification

Abstract Efficient delivery of biomolecules into neurons has significant impacts on therapeutic applications in the central nervous system (CNS) and fundamental neuroscience research. Existing viral and non‐viral delivery methods often suffer from inefficient intracellular access due to the endocyti...

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Main Authors: Xiaoqian Ge, Joseph B. Wekselblatt, Scott Elmore, Bo Wang, Tongtong Wang, Renjinming Dai, Tingting Zhang, Harsh Dave, Mohammadaref Ghaderi, Athul Raj Anilkumar, Bill Wang, Shashank R. Sirsi, Jung‐Mo Ahn, Mikhail G. Shapiro, Yuki Oka, Carlos Lois, Zhenpeng Qin
Format: Article
Language:English
Published: Wiley 2025-07-01
Series:Advanced Science
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Online Access:https://doi.org/10.1002/advs.202501033
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author Xiaoqian Ge
Joseph B. Wekselblatt
Scott Elmore
Bo Wang
Tongtong Wang
Renjinming Dai
Tingting Zhang
Harsh Dave
Mohammadaref Ghaderi
Athul Raj Anilkumar
Bill Wang
Shashank R. Sirsi
Jung‐Mo Ahn
Mikhail G. Shapiro
Yuki Oka
Carlos Lois
Zhenpeng Qin
author_facet Xiaoqian Ge
Joseph B. Wekselblatt
Scott Elmore
Bo Wang
Tongtong Wang
Renjinming Dai
Tingting Zhang
Harsh Dave
Mohammadaref Ghaderi
Athul Raj Anilkumar
Bill Wang
Shashank R. Sirsi
Jung‐Mo Ahn
Mikhail G. Shapiro
Yuki Oka
Carlos Lois
Zhenpeng Qin
author_sort Xiaoqian Ge
collection DOAJ
description Abstract Efficient delivery of biomolecules into neurons has significant impacts on therapeutic applications in the central nervous system (CNS) and fundamental neuroscience research. Existing viral and non‐viral delivery methods often suffer from inefficient intracellular access due to the endocytic pathway. Here, a neuron‐targeting and direct cytosolic delivery platform is discovered by using a 15‐amino‐acid peptide, termed the N1 peptide, which enables neuron‐specific targeting and cytosolic delivery of functional biomolecules. The N1 peptide initially binds hyaluronan in the extracellular matrix and subsequently passes the membrane of neurons without being trapped into endosome. This mechanism facilitates the efficient delivery of cell‐impermeable and photo‐stable fluorescent dye for super‐resolution imaging of dendritic spines, and functional proteins, such as Cre recombinase, for site‐specific genome modification. Importantly, the N1 peptide exhibits robust neuronal specificity across diverse species, including mice, rats, tree shrews, and zebra finches. Its targeting capability is further demonstrated through various administration routes, including intraparenchymal, intrathecal, and intravenous (i.v.) injections after blood‐brain barrier (BBB) opening with focused ultrasound (FUS). These findings establish the N1 peptide as a versatile and functional platform with significant potential for bioimaging and advanced therapeutic applications.
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spelling doaj-art-469bd6083db24ddeae52e5340d7a45802025-08-20T03:15:35ZengWileyAdvanced Science2198-38442025-07-011225n/an/a10.1002/advs.202501033In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome ModificationXiaoqian Ge0Joseph B. Wekselblatt1Scott Elmore2Bo Wang3Tongtong Wang4Renjinming Dai5Tingting Zhang6Harsh Dave7Mohammadaref Ghaderi8Athul Raj Anilkumar9Bill Wang10Shashank R. Sirsi11Jung‐Mo Ahn12Mikhail G. Shapiro13Yuki Oka14Carlos Lois15Zhenpeng Qin16Department of Biomedical Engineering University of Texas Southwestern Medical Center Dallas TX 75390 USADivision of Chemistry and Chemical Engineering California Institute of Technology Pasadena CA 91125 USADepartment of Chemistry and Biochemistry University of Texas at Dallas Richardson TX 75080‐3021 USADivision of Biology and Biological Engineering California Institute of Technology Pasadena CA 91125 USADivision of Biology and Biological Engineering California Institute of Technology Pasadena CA 91125 USADepartment of Bioengineering University of Texas at Dallas Richardson TX 75080‐3021 USADepartment of Mechanical Engineering University of Texas at Dallas Richardson TX 75080‐3021 USADepartment of Bioengineering University of Texas at Dallas Richardson TX 75080‐3021 USADepartment of Bioengineering University of Texas at Dallas Richardson TX 75080‐3021 USADepartment of Bioengineering University of Texas at Dallas Richardson TX 75080‐3021 USADepartment of Bioengineering University of Texas at Dallas Richardson TX 75080‐3021 USADepartment of Bioengineering University of Texas at Dallas Richardson TX 75080‐3021 USADepartment of Chemistry and Biochemistry University of Texas at Dallas Richardson TX 75080‐3021 USADivision of Chemistry and Chemical Engineering California Institute of Technology Pasadena CA 91125 USADivision of Biology and Biological Engineering California Institute of Technology Pasadena CA 91125 USADivision of Biology and Biological Engineering California Institute of Technology Pasadena CA 91125 USADepartment of Biomedical Engineering University of Texas Southwestern Medical Center Dallas TX 75390 USAAbstract Efficient delivery of biomolecules into neurons has significant impacts on therapeutic applications in the central nervous system (CNS) and fundamental neuroscience research. Existing viral and non‐viral delivery methods often suffer from inefficient intracellular access due to the endocytic pathway. Here, a neuron‐targeting and direct cytosolic delivery platform is discovered by using a 15‐amino‐acid peptide, termed the N1 peptide, which enables neuron‐specific targeting and cytosolic delivery of functional biomolecules. The N1 peptide initially binds hyaluronan in the extracellular matrix and subsequently passes the membrane of neurons without being trapped into endosome. This mechanism facilitates the efficient delivery of cell‐impermeable and photo‐stable fluorescent dye for super‐resolution imaging of dendritic spines, and functional proteins, such as Cre recombinase, for site‐specific genome modification. Importantly, the N1 peptide exhibits robust neuronal specificity across diverse species, including mice, rats, tree shrews, and zebra finches. Its targeting capability is further demonstrated through various administration routes, including intraparenchymal, intrathecal, and intravenous (i.v.) injections after blood‐brain barrier (BBB) opening with focused ultrasound (FUS). These findings establish the N1 peptide as a versatile and functional platform with significant potential for bioimaging and advanced therapeutic applications.https://doi.org/10.1002/advs.202501033across speciesgenome modificationneuron specific targetingpeptidessuper‐resolution imaging
spellingShingle Xiaoqian Ge
Joseph B. Wekselblatt
Scott Elmore
Bo Wang
Tongtong Wang
Renjinming Dai
Tingting Zhang
Harsh Dave
Mohammadaref Ghaderi
Athul Raj Anilkumar
Bill Wang
Shashank R. Sirsi
Jung‐Mo Ahn
Mikhail G. Shapiro
Yuki Oka
Carlos Lois
Zhenpeng Qin
In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification
Advanced Science
across species
genome modification
neuron specific targeting
peptides
super‐resolution imaging
title In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification
title_full In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification
title_fullStr In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification
title_full_unstemmed In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification
title_short In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification
title_sort in vivo cytosolic delivery of biomolecules into neurons for super resolution imaging and genome modification
topic across species
genome modification
neuron specific targeting
peptides
super‐resolution imaging
url https://doi.org/10.1002/advs.202501033
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