Microinjection of lentiviral vectors expressing small interfering RNAs directed against laminin receptor precursor mRNA prolongs the pre-clinical phase in scrapie-infected mice.
We examined therapeutic in vitro and in vivo approaches using lentivirus-based packaging of small interfering RNAs (siRNAs) targeting the non-integrin laminin receptor mRNA for treatment and prevention of prion disorders. Transfection of N2aSc(+) cells with recombinant plasmids expressing three different siRNAs, significantly reduced both the LRP (laminin receptor precursor) and PrP(Sc) levels by approximately 40-60 %. Stereotactic intracerebral microinjection of recombinant lentiviral vectors LVsiRNA-LRP 7 and 9 into the cortex of C57BL/6 wild-type mice resulted in a significant reduction of the LR levels in the cortex 15 days post-injection by 62 and 82 %, respectively. Intracerebral RML inoculation of C57BL/6 mice after microinjection with recombinant lentiviral vector LVsiRNA-LRP 7 into the hippocampus resulted in a significant reduction of both LRP and PrP(Sc) levels by 36 and 41 %, respectively, concomitant with a significant prolongation of the pre-clinical phase. Lentiviral vectors expressing siRNAs targeting LRP mRNA represent a novel delivery system for the treatment of transmissible spongiform encephalopathies.
Pflanz
Heike
H
Laboratorium für Molekulare Biologie - Genzentrum - Institut für Biochemie der LMU München, Feodor-Lynen Str. 25, D-81377 München, Germany.
Vana
Karen
K
Mitteregger
Gerda
G
Pace
Claudia
C
Messow
Diana
D
Sedlaczek
Cilli
C
Nikles
Daphne
D
Kretzschmar
Hans A
HA
Weiss
Stefan F T
SF
D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors; D006624:Hippocampus; D016086:Lentivirus / Q000235:genetics*; D008136:Longevity; D051379:Mice; D008810:Mice, Inbred C57BL; D008845:Microinjections; D012333:RNA, Messenger / Q000037:antagonists & inhibitors*; D034741:RNA, Small Interfering / Q000235:genetics*; D017474:Receptors, Laminin / Q000037:antagonists & inhibitors*; D012608:Scrapie / Q000517:prevention & control*; D016019:Survival Analysis; D013997:Time Factors
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D017474:Receptors, Laminin