Antisense delivery and protein knockdown within the intact central nervous system.
The ability to down regulate the expression of a specific protein within the intact central nervous system (CNS) is highly desirable from both a research and therapeutic perspective. Antisense has the potential to do this. However, problems of invasive antisense delivery methods and short half life of remain problematic. We overcome this by using Pluronic gel to provide a sustained delivery antisense oligodeoxynucleotides (ODN's) to the intact central nervous system and achieving rapid penetration throughout the spinal cord in 2-3 hours and significant knockdown of our target protein connexin 43 (Cx43) in 4-8 hours (recovering at 48-72 hours). Interestingly CY3-siRNA probes could not be detected penetrating the intact CNS and no knockdown the Cx43 was found. This approach with conventional ODNs could provide a faster and cheaper alternative to knockout mice in the investigation of the functions of specific proteins within the CNS and may also have therapeutic implications for drug discovery and development.
Cronin
Michael
M
Department of Anatomy and Developmental Biology, University College London, London, WC1E 6BT, United Kingdom.
Anderson
Patrick N
PN
Green
Colin R
CR
Becker
David L
DL
D000818:Animals; D018031:Connexin 43 / Q000096:biosynthesis* / Q000187:drug effects; D015536:Down-Regulation; D031541:Fluorescence Resonance Energy Transfer; D008297:Male; D020320:Oligoribonucleotides, Antisense / Q000008:administration & dosage* / Q000493:pharmacokinetics* / Q000502:physiology; D020442:Poloxamer; D034741:RNA, Small Interfering; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000737:chemistry / Q000378:metabolism*
D018031:Connexin 43; D020320:Oligoribonucleotides, Antisense; D034741:RNA, Small Interfering; D020442:Poloxamer