Gene silencing by synthetic U1 adaptors.
We describe a gene silencing method that employs a mechanism of action distinct from those of antisense and RNA interference. U1 Adaptors are bifunctional oligonucleotides with a 'target domain' complementary to a site in the target gene's terminal exon and a 'U1 domain' that binds to the U1 small nuclear RNA component of the U1 small nuclear ribonucleoprotein (U1 snRNP) splicing factor. Tethering of U1 snRNP to the target pre-mRNA inhibits poly(A)-tail addition, causing degradation of that RNA species in the nucleus. U1 Adaptors can inhibit both endogenous and reporter genes in a sequence-specific manner. Comparison of U1 Adaptors with small interfering RNA (siRNA) using a genome-wide microarray analysis indicates that U1 Adaptors have limited off-target effects and no detectable adverse effects on splicing. Further, targeting the same gene either with multiple U1 Adaptors or with a U1 Adaptor and siRNA strongly enhances gene silencing.
Goraczniak
Rafal
R
Rutgers University, 604 Allison Road, Piscataway, New Jersey 08854, USA.
Behlke
Mark A
MA
Gunderson
Samuel I
SI
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005786:Gene Expression Regulation; D020868:Gene Silencing; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D019908:Proto-Oncogene Proteins c-raf / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000235:genetics / Q000378:metabolism*; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014162:Transfection
D034741:RNA, Small Interfering; D012342:RNA, Small Nuclear; D017412:Ribonucleoprotein, U1 Small Nuclear; C405483:U1 small nuclear RNA; D008156:Luciferases; D019908:Proto-Oncogene Proteins c-raf