Fifty-one kilobase HSV-1 plasmid vector can be packaged using a helper virus-free system and supports expression in the rat brain.
Herpes simplex virus type 1 (HSV-1) plasmid vectors have a number of attractive features for gene transfer into neurons. In particular, the large size of the HSV-1 genome suggests that HSV-1 vectors might be designed to accommodate large inserts. We now report the construction and characterization of a 51 kb HSV-1 plasmid vector. This vector was efficiently packaged into HSV-1 particles using a helper virus-free packaging system. The structure of the packaged vector DNA was verified by both Southern blot and PCR analyses. A vector stock was microinjected into the rat striatum, the rats were sacrificed at 4 days after gene transfer, and numerous X-gal positive striatal cells were observed. This 51 kb vector was constructed using general principles that may support the routine construction of large vectors. Potential applications of such HSV-1 vectors include characterizing large promoter fragments or genomic clones and co-expressing multiple genes.
Wang
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X
Children's Hospital, Boston, MA, USA.
Zhang
G R
GR
Yang
T
T
Zhang
W
W
Geller
A I
AI
D000818:Animals; D001921:Brain / Q000378:metabolism*; D004279:DNA, Viral / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005800:Genes, Fungal; D005822:Genetic Vectors / Q000235:genetics*; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006651:Histocytochemistry; D007763:Lac Operon; D008297:Male; D008845:Microinjections; D009474:Neurons / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013238:Stereotaxic Techniques; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D004279:DNA, Viral; D001616:beta-Galactosidase