TY - JOUR
T1 - Structurally-discovered KLF4 variants accelerate and stabilize reprogramming to pluripotency
AU - Borisova, Evgeniia
AU - Nishimura, Ken
AU - An, Yuri
AU - Takami, Miho
AU - Li, Jingyue
AU - Song, Dan
AU - Matsuo-Takasaki, Mami
AU - Luijkx, Dorian
AU - Aizawa, Shiho
AU - Kuno, Akihiro
AU - Sugihara, Eiji
AU - Sato, Taka aki
AU - Yumoto, Fumiaki
AU - Terada, Tohru
AU - Hisatake, Koji
AU - Hayashi, Yohei
N1 - Publisher Copyright:
© 2021 The Author(s)
PY - 2022/1/21
Y1 - 2022/1/21
N2 - Non-genetically modified somatic cells can only be inefficiently and stochastically reprogrammed to pluripotency by exogenous expression of reprogramming factors. Low competence of natural reprogramming factors may prevent the majority of cells to successfully and synchronously reprogram. Here we screened DNA-interacting amino acid residues in the zinc-finger domain of KLF4 for enhanced reprogramming efficiency using alanine-substitution scanning methods. Identified KLF4 L507A mutant accelerated and stabilized reprogramming to pluripotency in both mouse and human somatic cells. By testing all the variants of L507 position, variants with smaller amino acid residues in the KLF4 L507 position showed higher reprogramming efficiency. L507A bound more to promoters or enhancers of pluripotency genes, such as KLF5, and drove gene expression of these genes during reprogramming. Molecular dynamics simulations predicted that L507A formed additional interactions with DNA. Our study demonstrates how modifications in amino acid residues of DNA-binding domains enable next-generation reprogramming technology with engineered reprogramming factors.
AB - Non-genetically modified somatic cells can only be inefficiently and stochastically reprogrammed to pluripotency by exogenous expression of reprogramming factors. Low competence of natural reprogramming factors may prevent the majority of cells to successfully and synchronously reprogram. Here we screened DNA-interacting amino acid residues in the zinc-finger domain of KLF4 for enhanced reprogramming efficiency using alanine-substitution scanning methods. Identified KLF4 L507A mutant accelerated and stabilized reprogramming to pluripotency in both mouse and human somatic cells. By testing all the variants of L507 position, variants with smaller amino acid residues in the KLF4 L507 position showed higher reprogramming efficiency. L507A bound more to promoters or enhancers of pluripotency genes, such as KLF5, and drove gene expression of these genes during reprogramming. Molecular dynamics simulations predicted that L507A formed additional interactions with DNA. Our study demonstrates how modifications in amino acid residues of DNA-binding domains enable next-generation reprogramming technology with engineered reprogramming factors.
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U2 - 10.1016/j.isci.2021.103525
DO - 10.1016/j.isci.2021.103525
M3 - Article
AN - SCOPUS:85122912155
SN - 2589-0042
VL - 25
JO - iScience
JF - iScience
IS - 1
M1 - 103525
ER -