Synergetic Targeted Delivery of Sleeping-Beauty Transposon System to Mesenchymal Stem Cells Using LPD Nanoparticles Modified with a Phage-Displayed Targeting Peptide.
Synergetic Targeted Delivery of Sleeping-Beauty Transposon System to Mesenchymal Stem Cells Using LPD Nanoparticles Modified with a Phage-Displayed Targeting Peptide.
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DOI：
10.1002/adfm.201102963
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发表时间：
2013-03-06
影响因子：
19
通讯作者：
Mao, Chuanbin
中科院分区：
文献类型：
--
作者：
Ma, Kun;Wang, Dong-Dong;Lin, Yiyang;Wang, Jianglin;Petrenko, Valery;Mao, Chuanbin
关键词：
有效的基因治疗的一个重要标准是有足够的染色体整合活性。睡美人(SB)转座子系统是一个允许转基因有效插入宿主基因组的质粒系统。然而，这种有效的插入只有在系统被递送到核之后才会发生。由于转座子不具有病毒载体的转导能力，因此该系统首先进入细胞然后进入细胞核的有效传递仍然是一个挑战。在这里，使用噬菌体展示技术使用主要的外套展示噬菌体文库来鉴定能够作为大鼠间充质干细胞(RMSCs)宿主的多肽(VTAMEPGQ)。一种被称为脂质体鱼精蛋白/DNA脂复合物(LPD)的纳米颗粒是由阳离子脂质体和鱼精蛋白、DNA和靶向多肽的阴离子络合物静电组装而成的。不同的多肽被包裹在LPD内，以提高其对rMSCs和核的靶向性。RMSC靶向多肽和核定位信号(NLS)多肽可以发挥协同作用，促进LPD的转导作用。归巢多肽指导LPD靶向MSCs，而NLS多肽指导转座子在LPD内化到细胞内后积聚到细胞核中，导致基因表达增加。这表明LPD内的RMSC靶向多肽和NLS多肽可以靶向rMSCs，并引导转座子进入细胞核。SB转座子进入细胞核后，可提高细胞染色体的插入率。靶向LPD对rMSCs没有明显的细胞毒性和分化潜能。因此，整合SB转座子和LPD系统是干细胞治疗中一种很有前途的非病毒基因传递载体。
An important criterion for effective gene therapy is sufficient chromosomal integration activity. The Sleeping Beauty (SB) transposon system is a plasmid system allowing efficient insertion of transgenes into the host genome. However, such efficient insertion occurs only after the system is delivered to nuclei. Since transposons do not have the transducing abilities of viral vectors, efficient delivery of this system first into cells and then into cell nuclei is still a challenge. Here, a phage display technique using a major coat displayed phage library is employed to identify a peptide (VTAMEPGQ) that can home to rat mesenchymal stem cells (rMSCs). A nanoparticle, called liposome protamine/DNA lipoplex (LPD), is electrostatically assembled from cationic liposomes and an anionic complex of protamine, DNA and targeting peptides. Various peptides are enveloped inside the LPD to improve its targeting capability for rMSCs and nuclei. The rMSC-targeting peptide and nuclear localization signal (NLS) peptide can execute the synergetic effect to promote transfection action of LPD. The homing peptide directs the LPD to target the MSCs, whereas the NLS peptide directs transposon to accumulate into nuclei once LPD is internalized inside the cells, leading to increased gene expression. This suggests that rMSC-targeting peptide and NLS peptide within LPD can target to rMSCs and then guide transposon into nuclei. After entering the nuclei, SB transposon increase the insertion rates into cellular chromosomes. The targeting LPD does not show obvious cell toxicity and influence on the differentiation potential of rMSCs. Therefore, the integration of SB transposon and LPD system is a promising nonviral gene delivery vector in stem cell therapy.
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影响因子：
10.8
作者：
Li, Shyh-Dar;Chono, Sumio;Huang, Leaf
通讯作者：
Huang, Leaf
DOI：
10.1196/annals.1348.001
发表时间：
2006-01-01
期刊：
OLIGONUCLEOTIDE THERAPEUTICS
影响因子：
--
作者：
Li, Shyh-Dar;Huang, Leaf
通讯作者：
Huang, Leaf
影响因子：
4.2
作者：
Sumiyoshi, Teiko;Holt, Nathalia G.;Kohn, Donald B.
通讯作者：
Kohn, Donald B.
影响因子：
15.8
作者：
Kelly, Kimberly A.;Bardeesy, Nabeel;Anbazhagan, Rajesh;Gurumurthy, Sushma;Berger, Justin;Alencar, Herlen;DePinho, Ronald A.;Mahmood, Umar;Weissleder, Ralph
通讯作者：
Weissleder, Ralph
影响因子：
14.8
作者：
Belur, Lalitha R.;Podetz-Pedersen, Kelly;McIvor, R. Scott
通讯作者：
McIvor, R. Scott