Gene delivery to postnatal rat brain by non-ventricular plasmid injection and electroporation.
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.
Molotkov
Dmitry A
DA
Neuroscience Center, University of Helsinki, Finland. dmitry.molotkov@helsinki.fi
Yukin
Alexey Y
AY
Afzalov
Ramil A
RA
Khiroug
Leonard S
LS
D000818:Animals; D001921:Brain / Q000502:physiology*; D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D008845:Microinjections / Q000379:methods*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D051381:Rats; D055647:Rats, Transgenic