文献检索文档翻译深度研究
Suppr Zotero 插件Zotero 插件
邀请有礼套餐&价格历史记录

新学期,新优惠

限时优惠:9月1日-9月22日

30天高级会员仅需29元

1天体验卡首发特惠仅需5.99元

了解详情
不再提醒
插件&应用
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
高级版
套餐订阅购买积分包
AI 工具
文献检索文档翻译深度研究
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利:CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2025

优化猪囊尾蚴转座子介导的基因转移方法在普通狨猴胚胎中。

Optimization of piggyBac transposon-mediated gene transfer method in common marmoset embryos.

机构信息

Laboratory of Applied Reproductive Science, Faculty of Agriculture, Shinshu University, Nagano, Japan.

Department of Neurophysiology, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan.

出版信息

PLoS One. 2023 Jun 9;18(6):e0287065. doi: 10.1371/journal.pone.0287065. eCollection 2023.


DOI:10.1371/journal.pone.0287065
PMID:37294815
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10256193/
Abstract

Generating non-human primate models of human diseases is important for the development of therapeutic strategies especially for neurodegenerative diseases. The common marmoset has attracted attention as a new experimental animal model, and many transgenic marmosets have been produced using lentiviral vector-mediated transgenesis. However, lentiviral vectors have a size limitation of up to 8 kb in length for transgene applications. Therefore, the present study aimed to optimize a piggyBac transposon-mediated gene transfer method in which transgenes longer than 8 kb were injected into the perivitelline space of marmoset embryos, followed by electroporation. We constructed a long piggyBac vector carrying the gene responsible for Alzheimer's disease. The optimal weight ratio of the piggyBac transgene vector to the piggyBac transposase mRNA was examined using mouse embryos. Transgene integration into the genome was confirmed in 70.7% of embryonic stem cells established from embryos injected with 1000 ng of transgene and transposase mRNA. Under these conditions, long transgenes were introduced into marmoset embryos. All embryos survived after transgene introduction treatment, and transgenes were detected in 70% of marmoset embryos. The transposon-mediated gene transfer method developed in this study can be applied to the genetic modification of non-human primates, as well as large animals.

摘要

生成人类疾病的非人类灵长类动物模型对于治疗策略的发展很重要,特别是对于神经退行性疾病。食蟹猴作为一种新的实验动物模型引起了人们的关注,许多转基因食蟹猴已经通过慢病毒载体介导的转基因技术产生。然而,慢病毒载体在转基因应用中长度限制为 8kb 以内。因此,本研究旨在优化一种基于 piggyBac 转座子的基因转移方法,该方法将长度超过 8kb 的转基因注入食蟹猴胚胎的卵周隙中,然后进行电穿孔。我们构建了一个携带阿尔茨海默病相关基因的长 piggyBac 载体。使用小鼠胚胎检查了 piggyBac 转基因载体与 piggyBac 转座酶 mRNA 的最佳重量比。用 1000ng 转基因和转座酶 mRNA 注射的胚胎建立的胚胎干细胞中,有 70.7%证实了转基因整合到基因组中。在这些条件下,长转基因被导入食蟹猴胚胎。转基因导入处理后所有胚胎均存活,70%的食蟹猴胚胎中检测到转基因。本研究开发的转座子介导的基因转移方法可应用于非人类灵长类动物和大动物的遗传修饰。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/383a/10256193/7dbf33620677/pone.0287065.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/383a/10256193/d99263de931b/pone.0287065.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/383a/10256193/f347b31c5f9b/pone.0287065.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/383a/10256193/7dbf33620677/pone.0287065.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/383a/10256193/d99263de931b/pone.0287065.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/383a/10256193/f347b31c5f9b/pone.0287065.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/383a/10256193/7dbf33620677/pone.0287065.g003.jpg

相似文献

[1]
Optimization of piggyBac transposon-mediated gene transfer method in common marmoset embryos.

PLoS One. 2023

[2]
Exogenous mRNA delivery and bioavailability in gene transfer mediated by piggyBac transposition.

BMC Biotechnol. 2013-9-26

[3]
Remobilizing deleted piggyBac vector post-integration for transgene stability in silkworm.

Mol Genet Genomics. 2015-6

[4]
Generation of non-viral, transgene-free hepatocyte like cells with piggyBac transposon.

Sci Rep. 2017-3-15

[5]
Generation of transgenic pigs by cytoplasmic injection of piggyBac transposase-based pmGENIE-3 plasmids.

Biol Reprod. 2014-5-8

[6]
Establishment of cell-based transposon-mediated transgenesis in cattle.

Theriogenology. 2016-4-15

[7]
piggyBac as a high-capacity transgenesis and gene-therapy vector in human cells and mice.

Dis Model Mech. 2013-3-8

[8]
PiggyBac transposon-mediated gene transfer in human cells.

Mol Ther. 2007-1

[9]
Vector modifications to eliminate transposase expression following piggyBac-mediated transgenesis.

Sci Rep. 2014-12-10

[10]
A piggyBac transposon- and gateway-enhanced system for efficient BAC transgenesis.

Dev Dyn. 2014-9

引用本文的文献

[1]
Functions and mechanisms of eukaryotic RNA-guided programmed DNA elimination.

Biochem Soc Trans. 2025-4-29

[2]
Non-viral generation of transgenic non-human primates via the piggyBac transposon system.

Nat Commun. 2025-3-24

[3]
Developing anti-TDE vaccine for sensitizing cancer cells to treatment and metastasis control.

NPJ Vaccines. 2025-1-27

本文引用的文献

[1]
Contemporary Transposon Tools: A Review and Guide through Mechanisms and Applications of , and for Genome Engineering.

Int J Mol Sci. 2021-5-11

[2]
Comparison of the effects of introducing the CRISPR/Cas9 system by microinjection and electroporation into porcine embryos at different stages.

BMC Res Notes. 2021-1-6

[3]
Efficient One-Step Knockout by Electroporation of Ribonucleoproteins Into Zona-Intact Bovine Embryos.

Front Genet. 2020-9-7

[4]
RNA Viruses as Tools in Gene Therapy and Vaccine Development.

Genes (Basel). 2019-3-1

[5]
Generation of transgenic marmosets using a tetracyclin-inducible transgene expression system as a neurodegenerative disease model.

Biol Reprod. 2017-11-1

[6]
Transgenic Monkey Model of the Polyglutamine Diseases Recapitulating Progressive Neurological Symptoms.

eNeuro. 2017-3-28

[7]
Delivery of Cas9 Protein into Mouse Zygotes through a Series of Electroporation Dramatically Increases the Efficiency of Model Creation.

J Genet Genomics. 2016-5-20

[8]
Efficient CRISPR/Cas9-Mediated Genome Editing in Mice by Zygote Electroporation of Nuclease.

Genetics. 2015-6

[9]
A hyperactive piggyBac transposon system is an easy-to-implement method for introducing foreign genes into mouse preimplantation embryos.

J Reprod Dev. 2015

[10]
The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies.

Gene Ther. 2015-2

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

推荐工具

医学文档翻译智能文献检索