PMC:7572125 / 17640-18743 JSONTXT

{"project":"2_test","denotations":[{"id":"33073769-16505370-27525623","span":{"begin":485,"end":489},"obj":"16505370"},{"id":"33073769-22000013-27525624","span":{"begin":598,"end":602},"obj":"22000013"},{"id":"33073769-26943900-27525625","span":{"begin":672,"end":676},"obj":"26943900"}],"text":"We replicated an experiment to test the microRNA dependency of the putative PTEN ceRNAs. This experiment used the same isogenic wild-type and DICER mutant (DicerEx5) HCT116 colon carcinoma cells as the original study. The DicerEx5 cell line, which was engineered to disrupt a well-conserved segment of the N-terminal helicase domain in exon 5 of DICER, while leaving the RNase III domains intact, displays a hypomorphic phenotype in the processing of mature microRNAs (Cummins et al., 2006). This experiment is similar to what was reported in Figure 3G–H and Supplemental Figure S3B of Tay et al., 2011 and described in Protocol 3 in the Registered Report (Phelps et al., 2016). Wild-type and DicerEx5 HCT116 cells were transfected with siRNAs targeting the same putative PTEN ceRNAs as the original study. Knockdown efficiency, measured by RT-qPCR, revealed the average reduction in gene expression relative to control siRNA was 81% in both cell lines for all putative PTEN ceRNAs, with the greatest biological variability in DicerEx5 HCT116 cells when targeting CNOT6L (Figure 3—figure supplement 1B)."}

{"project":"MyTest","denotations":[{"id":"33073769-16505370-27525623","span":{"begin":485,"end":489},"obj":"16505370"},{"id":"33073769-22000013-27525624","span":{"begin":598,"end":603},"obj":"22000013"},{"id":"33073769-26943900-27525625","span":{"begin":672,"end":676},"obj":"26943900"}],"namespaces":[{"prefix":"_base","uri":"https://www.uniprot.org/uniprot/testbase"},{"prefix":"UniProtKB","uri":"https://www.uniprot.org/uniprot/"},{"prefix":"uniprot","uri":"https://www.uniprot.org/uniprotkb/"}],"text":"We replicated an experiment to test the microRNA dependency of the putative PTEN ceRNAs. This experiment used the same isogenic wild-type and DICER mutant (DicerEx5) HCT116 colon carcinoma cells as the original study. The DicerEx5 cell line, which was engineered to disrupt a well-conserved segment of the N-terminal helicase domain in exon 5 of DICER, while leaving the RNase III domains intact, displays a hypomorphic phenotype in the processing of mature microRNAs (Cummins et al., 2006). This experiment is similar to what was reported in Figure 3G–H and Supplemental Figure S3B of Tay et al., 2011 and described in Protocol 3 in the Registered Report (Phelps et al., 2016). Wild-type and DicerEx5 HCT116 cells were transfected with siRNAs targeting the same putative PTEN ceRNAs as the original study. Knockdown efficiency, measured by RT-qPCR, revealed the average reduction in gene expression relative to control siRNA was 81% in both cell lines for all putative PTEN ceRNAs, with the greatest biological variability in DicerEx5 HCT116 cells when targeting CNOT6L (Figure 3—figure supplement 1B)."}