KLF4 N-terminal variance modulates induced reprogramming to pluripotency.
Bottom Line: Yet, subtle differences in methodology confound comparative studies of reprogramming mechanisms.Strikingly, global gene expression patterns elicited by published polycistronic cassettes diverged according to each KLF4 variant.Our data expose a Klf4 reference cDNA variation that alters polycistronic factor stoichiometry, predicts reprogramming hallmarks, and guides comparison of compatible public data sets.
Affiliation: Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 606-8507, Japan.Show MeSH
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Mentions: Considering the remarkable reprogramming phenotypes observed with polycistronic cassettes, we next addressed whether monocistronic expression of Klf4S or Klf4L in combination with polycistronic Oct3/4, Sox2, and c-Myc (OMS) had any detectable effect on iPSC derivation. OMS alone did not foster expansion of mCherry+ cells, which remained at a steady ∼6% through d18, nor did it activate SSEA-1 or Nanog-GFP expression (data not shown). Interestingly, expression of either Klf4 variant in combination with OMS resulted in nearly indistinguishable reprogramming phenotypes using the assays outlined in Figure 1 (see also Figures 2A, S2D, and S2E). These data demonstrate that the phenotypes associated with KLF4S and KLF4L arise mainly in the context of polycistronic factor linkage.
Affiliation: Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 606-8507, Japan.