KLF4 N-terminal variance modulates induced reprogramming to pluripotency.

Area of Science:

• Stem cell biology
• Molecular biology
• Epigenetics

Background:

• Induced pluripotency is typically achieved using OCT3/4, SOX2, KLF4, and c-MYC.
• Methodological variations hinder comparative studies of reprogramming mechanisms.

Purpose of the Study:

• To systematically assess cellular and molecular hallmarks of mouse somatic cell reprogramming using various polycistronic cassettes.
• To investigate the impact of KLF4 variants on reprogramming outcomes.

Main Methods:

• Utilized transposon-based delivery systems for polycistronic cassettes in mouse somatic cells.
• Analyzed reprogramming initiation, stabilization, protein stoichiometry, and global gene expression patterns.
• Compared reprogramming responses driven by different KLF4 variants.

Main Results:

• Reprogramming efficiency varied based on polycistronic cassette composition.
• A KLF4 variant differing by nine N-terminal amino acids resulted in dissimilar protein stoichiometry and reprogramming phenotypes.
• Supplementation or extension of the shorter KLF4 variant rescued protein levels and phenotypic disparities.
• Global gene expression patterns diverged significantly between KLF4 variants.

Conclusions:

• A specific KLF4 cDNA variation critically influences polycistronic factor stoichiometry and reprogramming hallmarks.
• Understanding KLF4 variant effects is essential for optimizing reprogramming protocols and comparing datasets.
• This study provides a reference for KLF4 variants in reprogramming studies.