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Updated: Jun 25, 2026
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RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells
Published on: November 26, 2018
piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells
Knut Woltjen1, Iacovos P Michael, Paria Mohseni
1Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario M5G 1X5, Canada.
Nature
|March 3, 2009
Summary
Efficient reprogramming of somatic cells into induced pluripotent stem (iPS) cells was achieved using the piggyBac (PB) transposition system. This virus-independent method allows for seamless removal of reprogramming factors, advancing cell-based therapies.
Area of Science:
- Cell Biology
- Genetics
- Biotechnology
Background:
- Four transcription factors (c-Myc, Klf4, Oct4, Sox2) induce pluripotency in somatic cells.
- Current methods like viral or plasmid transfection have limitations in efficiency and safety.
- The piggyBac (PB) transposition system offers a host-factor independent method for gene delivery.
Purpose of the Study:
- To demonstrate efficient reprogramming of fibroblasts into induced pluripotent stem (iPS) cells using the PB transposition system.
- To evaluate the ability to remove PB insertions from established iPS cell lines.
- To develop a simplified, virus-independent method for iPS cell production.
Main Methods:
- Delivery of doxycycline-inducible transcription factors via PB transposition into murine and human embryonic fibroblasts.
- Generation and characterization of stable iPS cell lines.
- Assessment of pluripotency markers and differentiation potential.
- Demonstration of seamless excision of PB insertions and reprogramming factors.
Main Results:
- Successful and efficient reprogramming of fibroblasts into iPS cells using PB transposition.
- Generated iPS cells expressed pluripotency markers and differentiated successfully.
- Demonstrated traceless removal of PB insertions and reprogramming factors from iPS lines.
- Achieved virus-independent simplification of iPS cell production.
Conclusions:
- PB transposition provides an efficient and versatile platform for iPS cell generation.
- The ability to remove PB elements is crucial for therapeutic applications.
- This simplified, virus-independent approach accelerates the field of reprogramming and cell-based therapies.
Related Concept Videos
01:29Somatic to iPS Cell Reprogramming
Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012 for this...
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Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore called induced pluripotent stem...
01:06Induced Pluripotent Stem Cells
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Somatic cells are...
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01:24Methods of Nuclear Reprogramming
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01:28Forced Transdifferentiation
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