Fast and Efficient Expression of Multiple Proteins in Avian Embryos Using mRNA Electroporation

被引：1

作者：

Tran, Martin ^{[1
]}

Dave, Mohit ^{[2
,3
]}

Lansford, Rusty ^{[1
,2
,3
]}

机构：

[1] Univ Southern Calif, Dept Biol Sci, Los Angeles, CA 90007 USA

[2] Childrens Hosp Los Angeles, Saban Res Inst, Dept Radiol & Dev Neurosci Program, Los Angeles, CA 90027 USA

[3] Univ Southern Calif, Keck Sch Med, Dept Radiol, Los Angeles, CA 90007 USA

来源：

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2019年 / 148期

关键词：

Developmental Biology; Issue; 148; mRNA electroporation; time-lapse microscopy; quail; avian; embryo; FRAP; GENE-EXPRESSION; XENOPUS-EMBRYOS; CHICK-EMBRYOS; CELLS; DYNAMICS;

D O I：

10.3791/59664

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

We report that mRNA electroporation permits fluorescent proteins to label cells in living quail embryos more quickly and broadly than DNA electroporation. The high transfection efficiency permits at least 4 distinct mRNAs to be co-transfected with similar to 87% efficiency. Most of the electroporated mRNAs are degraded during the first 2 h post-electroporation, permitting time-sensitive experiments to be carried out in the developing embryo. Finally, we describe how to dynamically image live embryos electroporated with mRNAs that encode various subcellular targeted fluorescent proteins.

引用

页数：13

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