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Lentiviral Mediated Delivery of shRNAs to hESCs and NPCs Using Low-cost Cationic Polymer Polyethylenimine (PEI)
Published on: May 24, 2022
Lentiviral vector-mediated gene delivery into human embryonic stem cells
Michal Gropp1, Benjamin Reubinoff
1Fodyn Savad Institute of Gene Therapy and Department of Bostetrics and Gynecology, Hadassah University Hospital, EinKerem, Jerusalem, Israel.
Methods in Enzymology
|December 13, 2006
Summary
Lentiviral vectors enable efficient genetic modification of human embryonic stem cells (hESCs), facilitating stable transgene expression and maintaining pluripotency for research applications.
Area of Science:
- Stem cell biology
- Gene therapy
- Molecular biology
Background:
- Human embryonic stem cells (hESCs) are pluripotent cells with potential for regenerative medicine and disease modeling.
- Effective genetic modification strategies are crucial for harnessing hESC potential in research.
- Lentiviral vectors offer a promising tool for gene delivery into hESCs.
Purpose of the Study:
- To review methods for lentiviral vector-mediated gene delivery into hESCs.
- To highlight the advantages of lentiviral vectors for hESC genetic modification.
- To discuss the stability of transgene expression and retention of pluripotency post-transduction.
Main Methods:
- Utilizing advanced, modified, replication-defective lentiviral vectors.
- Developing protocols for efficient transduction of hESCs.
- Culturing transduced hESCs under undifferentiated conditions and during differentiation.
Main Results:
- Lentiviral vectors efficiently transduce hESCs.
- Stable and high transgene expression is achieved in transduced hESCs.
- Transduced hESCs maintain self-renewal and pluripotent potential.
Conclusions:
- Lentiviral vectors are effective tools for the genetic modification of hESCs.
- Stable transgene expression and pluripotency are maintained, enabling further research.
- This technology is vital for advancing basic and applied research using hESCs.
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00:57Embryonic Stem Cells
Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
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00:58Embryonic Stem Cells
Embryonic stem (ES) cells are undifferentiated pluripotent cells, meaning they can produce any cell type in the body. This gives them tremendous potential in science and medicine since they can generate specific cell types for use in research or to replace body cells lost due to damage or disease.