1. Small interfering RNAs based on huntingtin trinucleotide repeats are highly toxic to cancer cells
Confluence over time of HeyA8 (top) or A549 (bottom) cells transfected with 1 nM of either siNT or the four TNR‐based siRNAs, siCAG/CUG, siCGG/CCG, siGAA/UUC, and siCGA/UCG. Values are mean ± SEM. n = 3/2 biological replicates (HeyA8/A549), eight technical replicates. Results of toxicity screens of 60 TNR‐based siRNAs in human HeyA8 (top) and mouse M565 (bottom) cells. Cells were reverse‐transfected in triplicate in 384‐well plates with 1 nM of each siRNA. In all cases, the complementary (sense) strand was inactivated by a 2′‐OMe modification in positions 1 and 2. Gray: no or less than 50% loss in viability; yellow: > 50% loss in viability; red: > 75% loss in viability. Family 7 is boxed in red as it is the only family in which all six duplexes were super toxic. Family 5 is boxed in blue. Its six members have the same GC content and the same nucleotide composition as the members of family 7. Family 10 is boxed in green. Each family contains three TNRs in the forward frame (for) and three TNRs that are the reverse complement (rev) of these TNRs. Values are mean ± SD. Three technical replicates. Correlation between the average of two viability screens performed in HeyA8 and two performed in M565 cells. The data points for the six members of TNR family 5 are labeled in blue those of family 7 are labeled in red. Pearson correlation and P‐value are given. Results of toxicity screen of 60 TNR‐based 6mer seeds in a nontoxic backbone siRNA (see Appendix Fig S5B) in HeyA8 cells. Values are mean ± SD. n = 3 technical replicates. Correlation between the average of two viability screens performed in HeyA8 with the 60 TNR‐based siRNAs and the screen performed with the 60 TNR‐based seeds. The data points for the six members of TNR family 7 are labeled in red and those of family 10 are labeled in green. Pearson correlation and P‐value are given.
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EMBO Rep, Volume: 19, Issue: 3, First published: 12 February 2018, DOI: ( /embr )