scholarly journals Imaging of the Entry Pathway of a Cell-Penetrating Peptide–DNA Complex From the Extracellular Space to Chloroplast Nucleoids Across Multiple Membranes in Arabidopsis Leaves

2021 ◽  
Vol 12 ◽  
Author(s):  
Kazusato Oikawa ◽  
Ayaka Tateishi ◽  
Masaki Odahara ◽  
Yutaka Kodama ◽  
Keiji Numata

Each plant cell has hundreds of copies of the chloroplast genome and chloroplast transgenes do not undergo silencing. Therefore, chloroplast transformation has many powerful potential agricultural and industrial applications. We previously succeeded in integrating exogenous genes into the chloroplast genome using peptide–DNA complexes composed of plasmid DNA and a fusion peptide consisting of a cell-penetrating peptide (CPP) and a chloroplast transit peptide (cpPD complex). However, how cpPD complexes are transported into the chloroplast from outside the cell remains unclear. Here, to characterize the route by which these cpPD complexes move into chloroplasts, we tracked their movement from the extracellular space to the chloroplast stroma using a fluorescent label and confocal laser scanning microscopy (CLSM). Upon infiltration of cpPD complexes into the extracellular space of Arabidopsis thaliana leaves, the complexes reached the chloroplast surface within 6h. The cpPD complexes reached were engulfed by the chloroplast outer envelope membrane and gradually integrated into the chloroplast. We detected several cpPD complexes localized around chloroplast nucleoids and observed the release of DNA from the cpPD. Our results thus define the route taken by the cpPD complexes for gene delivery from the extracellular space to the chloroplast stroma.

2014 ◽  
Vol 12 (36) ◽  
pp. 7036-7044 ◽  
Author(s):  
Oleksandr M. Zamotaiev ◽  
Viktoriia Y. Postupalenko ◽  
Volodymyr V. Shvadchak ◽  
Vasyl G. Pivovarenko ◽  
Andrey S. Klymchenko ◽  
...  

A new hydration-sensitive fluorescent label attached to the N-terminus of a cell-penetrating peptide allows visualization of the nanoscopic environment of its internalization pathway.


2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Makoto Oba ◽  
Takuma Kato ◽  
Kaori Furukawa ◽  
Masakazu Tanaka

2007 ◽  
Vol 18 (4) ◽  
pp. 1325-1331 ◽  
Author(s):  
Adams Amantana ◽  
Hong M. Moulton ◽  
Melissa L. Cate ◽  
Muralimohan T. Reddy ◽  
Tom Whitehead ◽  
...  

2011 ◽  
Vol 112 (9) ◽  
pp. 2454-2462
Author(s):  
Seung Won Yang ◽  
Seung-Min Lee ◽  
Eun Young Choi ◽  
Kyung Hye Lee ◽  
Soo Hyuk Kim ◽  
...  

Biomaterials ◽  
2014 ◽  
Vol 35 (13) ◽  
pp. 4082-4087 ◽  
Author(s):  
Likun Fei ◽  
Li-Peng Yap ◽  
Peter S. Conti ◽  
Wei-Chiang Shen ◽  
Jennica L. Zaro

2020 ◽  
Vol 17 ◽  
pp. 250-256
Author(s):  
Shanna J. Smith ◽  
Caroline M. Li ◽  
Robert G. Lingeman ◽  
Robert J. Hickey ◽  
Yilun Liu ◽  
...  

RSC Advances ◽  
2021 ◽  
Vol 11 (57) ◽  
pp. 36116-36124
Author(s):  
Omar Paulino da Silva Filho ◽  
Muhanad Ali ◽  
Rike Nabbefeld ◽  
Daniel Primavessy ◽  
Petra H. Bovee-Geurts ◽  
...  

Noncovalent functionalization with acylated cell-penetrating peptides achieves an efficient cellular uptake of PLGA and PEG-PLGA nanoparticles.


2008 ◽  
Vol 381 (5) ◽  
pp. 1133-1144 ◽  
Author(s):  
Yongchao Su ◽  
Rajeswari Mani ◽  
Tim Doherty ◽  
Alan J. Waring ◽  
Mei Hong

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