Description

The RESC (RNAi Escape Strategy Construct) strategy provides a simple and efficient method to validate RNAi-mediated gene knockdown, rule out off-target effects, or perform protein structure-function analysis. In principle, frequent silent mismatches in the complete RNAi target sequence allow expression of a wild type rescue protein through exploiting the degenerate genetic code. In species like Drosophila RNAi induced knockdown is not transitive and therefore non-targeted sequence of the targzeted gene may remain wild type.

For generation of a RESC sequence containing a maximum number of silent mutations the forward DNA sequence of the target region has to be pasted into the DNA sequence window below. Importantly, the first nucleotide of the pasted sequence must be the first nucleotide of a codon.
The sequences of oligonucleotides primers that can be used forstandard PCR are provided based on the maximum primer length andoverlap between the primers.

Design your RNAi rescue construct

 

Enter your DNA sequence here:

Start position of target region (is start of a codon)

End position of target region

Maximum length of oligonucleotides

Preferred overlap between oligo's

Citing RESC

Schulz JG, David G, Hassan BA. A novel method for tissue-specific RNAi rescue in Drosophila.Nucleic Acids Res. 2009 Jul;37(13):e93

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