shRNA/miRNAValidationServiceviaCell-basedReporterSystem:
-CloningORFofgeneofinterests(GOI)intoarAAVcisvectorwithareporter.
-Designandsynthesize~6shRNAoligoesfollowedbycloningthemtotheaboverAAVcisconstructtoobtainshRNAvalidationplasmids.
-FormiRNAvalidation,synthesizepre-miRNAoligoesfollowedbycloningthemtotheaboverAAVcisconstructtoobtainshRNAvalidationplasmid.
-TransienttransfectiontheshRNA/miRNAvalidationplasmids.
-QuantificationoftargetgenesilencingviaFACS(forGFPexpression)orluminometer(forluciferaseexpression).
AvailableshRNA/miRNAValidationPlatforms:
-ConstitutiveshRNAknockdown
-InducIBLeshRNAknockdown
GuaranteedKnockdown:
Weguaranteeatleast1(one)shRNAwith>80%knockdownofmRNAlevelinthetestedcellsbyscreening~6shRNA/miRNAoligoes.
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Figure1. ACartoonShowingshRNAValidationProcessforshRNArAAVProduction.