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Peer-reviewedCC BY-NC-NDBlood science (Baltimore, Md.)

Single-episome delivery of BOSMK with miR-142-gated KLF4 streamlines PBMC reprogramming to iPSCs.

  • Xiang Li1,2
  • Wei Wen1,2
  • Jian-Qiao Chang1,2
  • Feng Zhang1,2
  • Zhu-Ying Gao1,2
  • Shu-Man Zhang1,2
  • Guo-Hua Li1,2
  • Bin Liu1,2
  • Jue Wang1,2
  • Hang Li1,2
  • Chong-Yan Cheng1,2
  • Li-Ming Zhang1,2
2 more authors
  • Jian-Ping Zhang1,2
  • Xiao-Bing Zhang1,2
  1. 1State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.
  2. 2Tianjin Institutes of Health Science, Tianjin 301600, China.
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Abstract

Peripheral blood-derived induced pluripotent stem cells (iPSCs) are attractive for clinical applications, yet multi-plasmid episomal protocols reduce delivery completeness and complicate good manufacturing practice (GMP) workflows. Here, we assembled OCT4, SOX2, MYC, BCL-XL, and KLF4 into a single OriP/EBNA1 episome and tuned KLF4 expression by placing it under the elongation factor 1 alpha (EF1α) promoter with 2 tandem miR-142-3p target sites in its 3'UTR. Across donors, the single-episome system produced robust day-14 colony yields normalized per 1 × 105 input cells. A green fluorescent protein (GFP) reporter demonstrated hematopoietic miR-142-dependent suppression, and the miR-gated construct exhibited significantly faster EBNA1 sequence loss by day 4. Resulting iPSC lines expressed TRA-1-60/TRA-1-81, SSEA4, NANOG, and OCT4, maintained normal karyotypes, and generated derivatives of all three germ layers. This miR-142-gated EF1α-KLF4 single-episome streamlines peripheral blood mononuclear cells (PBMCs) reprogramming and provides a GMP-amenable, non-integrating route to high-quality iPSCs.

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Cite

BibTeXRIS
BibTeX
@article{li2026singleepisome,
  title = {{Single-episome delivery of BOSMK with miR-142-gated KLF4 streamlines PBMC reprogramming to iPSCs.}},
  author = {Xiang Li and Wei Wen and Jian-Qiao Chang and Feng Zhang and Zhu-Ying Gao and Shu-Man Zhang and Guo-Hua Li and Bin Liu and Jue Wang and Hang Li and Chong-Yan Cheng and Li-Ming Zhang and Jian-Ping Zhang and Xiao-Bing Zhang},
  journal = {Blood science (Baltimore, Md.)},
  year = {2026},
  month = nov,
  doi = {10.1097/bs9.0000000000000277},
  url = {https://europepmc.org/article/MED/42997867},
  note = {CC BY-NC-ND},
}
RIS
TY  - JOUR
TI  - Single-episome delivery of BOSMK with miR-142-gated KLF4 streamlines PBMC reprogramming to iPSCs.
AU  - Xiang Li
AU  - Wei Wen
AU  - Jian-Qiao Chang
AU  - Feng Zhang
AU  - Zhu-Ying Gao
AU  - Shu-Man Zhang
AU  - Guo-Hua Li
AU  - Bin Liu
AU  - Jue Wang
AU  - Hang Li
AU  - Chong-Yan Cheng
AU  - Li-Ming Zhang
AU  - Jian-Ping Zhang
AU  - Xiao-Bing Zhang
PY  - 2026
DA  - 2026/11/05
JO  - Blood science (Baltimore, Md.)
DO  - 10.1097/bs9.0000000000000277
UR  - https://europepmc.org/article/MED/42997867
AB  - Peripheral blood-derived induced pluripotent stem cells (iPSCs) are attractive for clinical applications, yet multi-plasmid episomal protocols reduce delivery completeness and complicate good manufacturing practice (GMP) workflows. Here, we assembled OCT4, SOX2, MYC, BCL-XL, and KLF4 into a single OriP/EBNA1 episome and tuned KLF4 expression by placing it under the elongation factor 1 alpha (EF1α) promoter with 2 tandem miR-142-3p target sites in its 3'UTR. Across donors, the single-episome system produced robust day-14 colony yields normalized per 1 × 105 input cells. A green fluorescent protein (GFP) reporter demonstrated hematopoietic miR-142-dependent suppression, and the miR-gated construct exhibited significantly faster EBNA1 sequence loss by day 4. Resulting iPSC lines expressed TRA-1-60/TRA-1-81, SSEA4, NANOG, and OCT4, maintained normal karyotypes, and generated derivatives of all three germ layers. This miR-142-gated EF1α-KLF4 single-episome streamlines peripheral blood mononuclear cells (PBMCs) reprogramming and provides a GMP-amenable, non-integrating route to high-quality iPSCs.
SN  - 2543-6368
ER  -