Gene Delivery to Postnatal Rat Brain by Non-ventricular Plasmid Injection and Electroporation
Overview
Affiliations
Creation of transgenic animals is a standard approach in studying functions of a gene of interest in vivo. However, many knockout or transgenic animals are not viable in those cases where the modified gene is expressed or deleted in the whole organism. Moreover, a variety of compensatory mechanisms often make it difficult to interpret the results. The compensatory effects can be alleviated by either timing the gene expression or limiting the amount of transfected cells. The method of postnatal non-ventricular microinjection and in vivo electroporation allows targeted delivery of genes, siRNA or dye molecules directly to a small region of interest in the newborn rodent brain. In contrast to conventional ventricular injection technique, this method allows transfection of non-migratory cell types. Animals transfected by means of the method described here can be used, for example, for two-photon in vivo imaging or in electrophysiological experiments on acute brain slices.
Recent Advances in In Vivo Somatic Cell Gene Modification in Newborn Pups.
Nakamura S, Morohoshi K, Inada E, Sato Y, Watanabe S, Saitoh I Int J Mol Sci. 2023; 24(20).
PMID: 37894981 PMC: 10607593. DOI: 10.3390/ijms242015301.
Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation.
Yu C, Stefanson O, Liu Y, Wang Z J Vis Exp. 2018; (135).
PMID: 29782018 PMC: 6101102. DOI: 10.3791/57649.
In vivo rapid gene delivery into postmitotic neocortical neurons using iontoporation.
De la Rossa A, Jabaudon D Nat Protoc. 2014; 10(1):25-32.
PMID: 25474030 DOI: 10.1038/nprot.2015.001.
Microinjection of membrane-impermeable molecules into single neural stem cells in brain tissue.
Wong F, Haffner C, Huttner W, Taverna E Nat Protoc. 2014; 9(5):1170-82.
PMID: 24762784 DOI: 10.1038/nprot.2014.074.
Broderick K, Chan A, Lin F, Shen X, Kichaev G, Khan A Mol Ther Nucleic Acids. 2013; 1:e11.
PMID: 23344722 PMC: 3381603. DOI: 10.1038/mtna.2012.1.