A gene delivery system containing nuclear localization signal: Increased nucleus import and transfection efficiency with the assistance of RanGAP1

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• 作者：Kang Chen^a ; ¹ ; Lingling Guo^b ; ¹ ; Jiulong Zhang^a ; Qing Chen^a ; Kuanglei Wang^a ; Chenxi Li^a ; Weinan Li^a ; Mingxi Qiao^a ; Xiuli Zhao^a ; Haiyang Hu^a ; ^{haiyang_hu@hotmail.com} ; Dawei Chen^a ; ^c ; ^{chendawei@syphu.edu.cn}
• 关键词：PAMAM-OH ; hydroxyl terminal PAMAM dendrimer (G4) ; PAMAM ; amino terminal PAMAM dendrimer (G4) ; PAMS ; SMLC-modified PAMAM-OH ; PAMN ; L-Norvaline-modified PAMAM-OH. RanGAP1 ; GTPase-activating protein 1 ; RanGAP1 cells ; KB cells overexpressing RanGAP1 ; G1 cells ; KB cells in G1 phase ; G2/M cells ; KB cells in G2/M phase
• 刊名：Acta Biomaterialia
• 出版年：2017
• 出版时间：15 January 2017
• 年：2017
• 卷：48
• 期：Complete
• 页码：215-226
• 全文大小：2341 K
• 卷排序：48

文摘

In the present report, a degradable gene delivery system (PAMS/DNA/10NLS) containing nucleus location signal peptide (NLS) was prepared. The agarose gel electrophoresis, particle size and zeta potential of PAMS/DNA/10NLS were similar to those of PAMS/DNA, which proved that NLS did not affect the interaction between PAMS and DNA. PAMS/DNA/10NLS exhibited marked extracellular and intracellular degradation under acidic conditions. The degradation was believed to allow NLS to come into contact with importins easily, which was able to mediate the nucleus import. With the help of NLS, PAMS/DNA/10NLS exhibited a higher transfection capability than PAMS/DNA. Moreover, the transfection of PAMS/DNA/10NLS was less dependent on the breakdown of the nucleus envelope than PAMS/DNA. Considering that GTPase-activating protein 1 (RanGAP1) was able to activate the endogenous GTPase, which was necessary for NLS-mediated nucleus import, RanGAP1 overexpressed cells (RanGAP1 cells) were produced. This result showed that RanGAP1 cells had higher GTPase activities than normal cells. Both the nucleus import and transfection efficiency of PAMS/DNA/10NLS were markedly higher in RanGAP1 cells than that in normal cells. The in vivo transfection results also showed that the transfection efficiency of PAMS/DNA/10NLS was higher in RanGAP1 pre-treated mice than that in normal mice. These findings showed that PAMS/DNA/10NLS is a promising gene delivery system with the assistance of RanGAP1.Statement of SignificanceThe present report describes the increased transfection efficiency of a degradable gene delivery system (PAMS/DNA/10NLS) containing nuclear location signal (NLS) with the assistance of GTPase-activating protein 1 (RanGAP1). The physicochemical properties of PAMS/DNA/10NLS were similar to those of PAMS/DNA. PAMS/DNA/10NLS exhibited great extracellular and intracellular degradations, which might allow NLS to contact with importins easily. With the help of NLS, PAMS/DNA/10NLS exhibited a higher transfection capability than PAMS/DNA. The transfection of PAMS/DNA/10NLS had less dependence on the breakdown of nuclear envelope. Both the nuclear import and transfection efficiency of PAMS/DNA/10NLS were higher in RanGAP1 overexpressed cells than that in normal cells. Moreover, the transfection efficiency of PAMS/DNA/10NLS was higher in RanGAP1 pre-treated mice than that in normal mice.