KLF4 N-terminal variance modulates induced reprogramming to pluripotency

Stem Cell Reports
Shin-Il KimKnut Woltjen

Abstract

As the quintessential reprogramming model, OCT3/4, SOX2, KLF4, and c-MYC re-wire somatic cells to achieve induced pluripotency. Yet, subtle differences in methodology confound comparative studies of reprogramming mechanisms. Employing transposons, we systematically assessed cellular and molecular hallmarks of mouse somatic cell reprogramming by various polycistronic cassettes. Reprogramming responses varied in the extent of initiation and stabilization of transgene-independent pluripotency. Notably, the cassettes employed one of two KLF4 variants, differing only by nine N-terminal amino acids, which generated dissimilar protein stoichiometry. Extending the shorter variant by nine N-terminal amino acids or augmenting stoichiometry by KLF4 supplementation rescued both protein levels and phenotypic disparities, implicating a threshold in determining reprogramming outcomes. 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.

References

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Citations

Feb 18, 2016·Nature Reviews. Molecular Cell Biology·Kazutoshi Takahashi, Shinya Yamanaka
Jan 16, 2016·Stem Cells International·Clair E WeidgangMartin Mueller
Nov 22, 2015·Seminars in Immunology·Peter KaragiannisShin Kaneko
Jul 28, 2015·Trends in Biotechnology·Lauren E Woodard, Matthew H Wilson
Aug 19, 2015·Stem Cell Reports·Eleni ChantzouraKeisuke Kaji
Oct 18, 2018·Physiological Reviews·Peter KaragiannisKenji Osafune
Jun 22, 2016·Oncotarget·Jacqueline FerralliMartin Degen
Apr 26, 2016·Development, Growth & Differentiation·Lihi Ben-Reuven, Orly Reiner
Oct 8, 2015·Development·Kazutoshi Takahashi, Shinya Yamanaka
Apr 13, 2018·Nature Communications·Takashi IkedaShinji Masui
Nov 14, 2019·Cell Reports·Phuong Linh BuiKoji Hisatake
Jan 15, 2019·Stem Cell Reports·Harunobu KagawaKnut Woltjen

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Datasets Mentioned

BETA
U70662.1
and
U20344.1
AAC52939.1
AAC04892.1

Methods Mentioned

BETA
fluorescence-activated cell sorting
transfection
FACS
transfections
transgenic
PCR
genotyping
Fluorescence

Software Mentioned

Tree Star
FlowJo
Sequencher
GeneSpring GX
GeneSpring
- Quant
GeneCodes
BD FACSDiva
CL

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