Targeted sequence alteration of a chromosomal locus in mouse liver.
Targeted sequence alteration would be an attractive method in gene therapy and biotechnology. To achieve in vivo targeted sequence alteration, a tailed duplex DNA consisting of annealed 35mer and 794mer single-stranded DNAs was delivered by means of hydrodynamic tail vein injection into liver of transgenic mouse harboring a reporter gene (the rpsL gene) in its genome. The tailed DNA was designed for a conversion of ATC to AGC at codon 80 of the rpsL transgene. The anticipated T-->G sequence alteration was induced in the transgene in the liver with an efficiency of approximately 0.1%. These results demonstrate the significant potential of this method for applications in gene therapy and biotechnology.
Kamiya
Hiroyuki
H
Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan. hirokam@pharm.hokudai.ac.jp
Uchiyama
Masayuki
M
Piao
Jingshu
J
Nakatsu
Yoshimichi
Y
Tsuzuki
Teruhisa
T
Harashima
Hideyoshi
H
D000818:Animals; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods; D033481:Chromosomes, Mammalian / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000008:administration & dosage*; D005260:Female; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D016678:Genome; D008099:Liver / Q000378:metabolism*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D017422:Sequence Analysis, DNA
D004277:DNA, Single-Stranded