Allele-specific gene silencing by RNA interference (RNAi) is therapeutically useful for specifically suppressing the expression of alleles associated with disease. To realize such allele-specific RNAi (ASPRNAi), the design and assessment of small interfering RNA (siRNA) duplexes conferring ASP-RNAi is vital, but is also difficult. Here, we show ASP-RNAi against the Swedish- and London-type amyloid precursor protein (APP) variants related to familial Alzheimer's disease using two reporter alleles encoding the Photinus and Renilla luciferase genes and carrying mutant and wild-type allelic sequences in their 3'-untranslated regions. We examined the effects of siRNA duplexes against the mutant alleles in allele-specific gene silencing and off-target silencing against the wild-type allele under heterozygous conditions, which were generated by cotransfecting the reporter alleles and siRNA duplexes into cultured human cells. Consistently, the siRNA duplexes determined to confer ASP-RNAi also inhibited the expression of the bona fide mutant APP and the production of either amyloid beta 40- or 42-peptide in Cos-7 cells expressing both the full-length Swedish- and wild-type APP alleles. The present data suggest that the system with reporter alleles may permit the preclinical assessment of siRNA duplexes conferring ASP-RNAi, and thus contribute to the design and selection of the most suitable of such siRNA duplexes.
Allele - specific gene silencing by RNA interference ( RNAi ) is therapeutically useful for specifically suppressing the expression of alleles associated with disease . To realize such allele - specific RNAi ( ASPRNAi ) , the design and assessment of small interfering RNA ( siRNA ) duplexes conferring ASP - RNAi is vital , but is also difficult . Here , we show ASP - RNAi against the Swedish- and London - type amyloid precursor protein ( APP ) variants related to familial Alzheimer 's disease using two reporter alleles encoding the Photinus and Renilla luciferase genes and carrying mutant and wild - type allelic sequences in their 3'-untranslated regions . We examined the effects of siRNA duplexes against the mutant alleles in allele - specific gene silencing and off - target silencing against the wild - type allele under heterozygous conditions , which were generated by cotransfecting the reporter alleles and siRNA duplexes into cultured human cells . Consistently , the siRNA duplexes determined to confer ASP - RNAi also inhibited the expression of the bona fide mutant APP and the production of either amyloid beta 40- or 42-peptide in Cos-7 cells expressing both the full - length Swedish- and wild - type APP alleles . The present data suggest that the system with reporter alleles may permit the preclinical assessment of siRNA duplexes conferring ASP - RNAi , and thus contribute to the design and selection of the most suitable of such siRNA duplexes .
Which disease the London mutation involved in?
Which disease the London mutation involved in ?
Alzheimer's Disease
Alzheimer's Disease AD