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In vivo Reprogramming of Adult Somatic Cells to Pluripotency by Overexpression of Yamanaka Factors
Published on: December 17, 2013
Reprogramming of murine and human somatic cells using a single polycistronic vector.
Bryce W Carey1, Styliani Markoulaki, Jacob Hanna
1Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.
Summary
This study introduces a single-virus method for generating induced pluripotent stem cells (iPSCs) using 2A peptides. This approach reduces viral integration risks, enabling efficient reprogramming of mouse and human somatic cells.
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Genetic Engineering
Background:
- Induced pluripotent stem cells (iPSCs) offer patient-specific regenerative medicine potential.
- Viral delivery of reprogramming factors poses risks like insertional mutagenesis.
- Current methods require multiple viral vectors for efficient iPSC generation.
Purpose of the Study:
- To develop a safer and more efficient method for generating iPSCs.
- To reduce the number of viral vectors needed for somatic cell reprogramming.
- To investigate the efficacy of polycistronic expression for iPSC generation.
Main Methods:
- Utilized 2A "self-cleaving" peptides for polycistronic expression of reprogramming factors (Oct4, Sox2, Klf4, c-Myc) from a single viral vector.
- Applied the single-virus system to reprogram both embryonic and adult somatic mouse cells.
- Generated human induced pluripotent stem (hiPS) cell lines from human keratinocytes.
Main Results:
- Successfully generated iPSCs from mouse embryonic fibroblasts using a single proviral copy.
- Demonstrated efficient reprogramming of both embryonic and adult somatic mouse cells.
- Established human iPS cell lines from keratinocytes, confirming the method's applicability to human cells.
Conclusions:
- A single polycistronic virus effectively reprograms somatic cells into iPSCs, minimizing insertional mutagenesis risks.
- This novel approach enhances the safety and efficiency of iPSC generation for therapeutic applications.
- The method is validated in both mouse and human somatic cells, paving the way for clinical translation.
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01:24Methods of Nuclear Reprogramming
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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...