Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys(2)-His(2) Zinc-finger Domains

doi:10.3791/52814

	Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys(2)-His(2) Zinc-finger Domains
	Gaj, Thomas 1,2; Liu, Jia1,2,3
	2015-03
发表期刊	JOVE-JOURNAL OF VISUALIZED EXPERIMENTS (IF:1.2[JCR-2023],1.4[5-Year])
ISSN	1940-087X
期号	97
发表状态	已发表
DOI	10.3791/52814
摘要	Due to their modularity and ability to be reprogrammed to recognize a wide range of DNA sequences, Cys(2)-His(2) zinc-finger DNA-binding domains have emerged as useful tools for targeted genome engineering. Like many other DNA-binding proteins, zinc-fingers also possess the innate ability to cross cell membranes. We recently demonstrated that this intrinsic cell-permeability could be leveraged for intracellular protein delivery. Genetic fusion of zinc-finger motifs leads to efficient transport of protein and enzyme cargo into a broad range of mammalian cell types. Unlike other protein transduction technologies, delivery via zinc-finger domains does not inhibit enzyme activity and leads to high levels of cytosolic delivery. Here a detailed step-by-step protocol is presented for the implementation of zinc-finger technology for protein delivery into mammalian cells. Key steps for achieving high levels of intracellular zinc-finger-mediated delivery are highlighted and strategies for maximizing the performance of this system are discussed.
关键词	Molecular Biology Issue 97 protein delivery cell-penetrating peptide zinc-finger protein transduction domain chemical biology molecular biology
收录类别	SCI
语种	英语
资助项目	National Institutes of Health[DP1CA174426]
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000361534300074
出版者	JOURNAL OF VISUALIZED EXPERIMENTS
WOS关键词	FUNCTIONAL PROTEINS ; MINIATURE PROTEINS ; MEDIATED DELIVERY ; IN-VITRO ; PEPTIDE ; TRANSDUCTION
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/2245
专题	免疫化学研究所_特聘教授组_抗体化学实验室免疫化学研究所_PI研究组_刘佳组
通讯作者	Gaj, Thomas
作者单位	1.Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA 2.Scripps Res Inst, Dept Cell & Mol Biol, La Jolla, CA USA 3.ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai, Peoples R China
推荐引用方式 GB/T 7714	Gaj, Thomas,Liu, Jia. Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys(2)-His(2) Zinc-finger Domains[J]. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS,2015(97).
APA	Gaj, Thomas,&Liu, Jia.(2015).Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys(2)-His(2) Zinc-finger Domains.JOVE-JOURNAL OF VISUALIZED EXPERIMENTS(97).
MLA	Gaj, Thomas,et al."Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys(2)-His(2) Zinc-finger Domains".JOVE-JOURNAL OF VISUALIZED EXPERIMENTS .97(2015).