Gene delivery methods |
Viral delivery system |
Lentivirus |
Advantages |
- Infect both dividing and non-dividing cells |
- Recombinant can be pseudotyped to broaden tropism |
- Highly efficient |
- Constitutive and inducible expression can be attained |
Disadvantage |
- Integration into host genome |
Retrovirus |
Advantages |
- Undergo silencing in ES cells |
- Highly efficient |
Disadvantage |
- Integration into host genome |
Adenovirus |
Advantage |
- Non-integrating vector |
Disadvantages |
- Poor gene transfer |
- Control of gene expression level difficult |
Non-viral system |
Episomal plasmids |
Advantages |
- Non-integrating vector |
- Able to replicate autonomously giving rise to prolonged expression |
Disadvantage |
- Low reprogramming efficiency |
Excision strategies |
Cre-loxP recombination |
Advantages |
- Gene expression profile closer to hES cells compared to human iPS cells |
- Efficient excision of integrated transgenes |
Disadvantages |
- Genomic instability and genome rearrangements |
- loxP site remain integrated in genome |
piggyBac transposon |
Advantages |
- Self-excision |
- Precise and efficient excision |
Disadvantage |
- pBt gene may remain active post-transposition |
Non-DNA methods |
Protein-mediated |
Poly-arginine peptide tags |
Advantages |
- Reprogrammed cells from embryonic fibroblast cells were indistinguishable from the classic embryonic stem cells |
Disadvantages |
- Not clear is can be used for adult cells |
- Short half-life |
Manipulation of cell-culture conditions |
Oxygen level manipulation |
Advantages |
- No genetic manipulation |
- No protein carriers needed |
Disadvantage |
- Short-term induction |
miRNAs |
miR-294, miR-302 |
Advantages |
- Able to replace c-Myc |
- Increased efficiency |
Disadvantages |
- Not specific |
- Target point may be too upstream in the cell signalling pathway |
- Expression not easily controlled post-induction |
Enhancing DNA reprogramming efficiencies |
Small molecules and soluble factors |
Chromatin modification modifiers |
Advantages |
- Modulate chromatin modification |
- Target cell-signalling pathways |
Disadvantage |
- Non-specific and broad action |
- May cause dysregulation of gene expression |