Creative Bioarray is a leading provider of cell reprogramming technology services. We offer a wide range of cell reprogramming services and help our customers improve the efficiency of cell reprogramming by developing simpler gene delivery/expression systems. The cell reprogramming-related biotechnology services we provided can fully meet the research needs of our customers.
Background
The technology of iPSCs reprogramming has great potential to enable medical applications of patient-derived engineered stem cells. iPSCs were originally established by ectopic expression of multiple TFs using lentiviral or retroviral vectors. But these genomic integration methods may generate tumorigenic insertional mutations, and silencing and reactivation of integrated reprogramming genes may affect the in vitro differentiation capacity of iPSCs and the functionality of iPS-derived cells.
To avoid permanent genetic modifications of newly reprogrammed iPSCs to meet the stringent safety requirements for clinical treatment, new platforms are needed to reprogram cells into iPSCs without the use of recombinant DNA or viral vectors. Therefore, researchers have focused on developing new methods to establish iPSCs that do not carry exogenous genetic materials. Currently, various approaches were explored to derive freely transgenic iPSCs, such as small-loop DNA vectors, plasmid vectors, piggyBac, mRNA, proteins, adenovirus, Sendai virus, and small molecules.
Fig.1 Structure of a novel replication-defective and persistent Sendai virus vector. (Nishimura, 2017)
Our Strategies
In order to efficiently deliver exogenous factors into various mammalian somatic cells, Creative Bioarray has developed various strategies to create simpler gene delivery/expression systems suitable for cell reprogramming. Our strategies include, but are not limited to:
- Sendai virus
- Efficient and harmless
- Simultaneous delivery of multiple exogenous genes
- Generation of non-integrated and feeder-free iPSCs
- mRNA
- Efficient and rapid
- Integration-free and footprint-free
- Cost-effective
- RNA can be easily and selectively eliminated
- Episomal plasmids
- High transfection efficiency
- Wide application to a large number of somatic cell types
- Long transgene ectopic expression
Sendai virus is a non-fragmented negative strand RNA virus that neither integrates into the host genome nor alters the genetic information of the host cell. Moreover, using Sendai viruses as gene delivery/expression systems can infect a wide range of cell types in quiescent and proliferative states with high transduction efficiency. Advantages of this strategy include:
The use of mRNA to express reprogramming factors provides a method for preparing transgene-free iPSCs. A single RNA strand contains multiple reprogramming factors and does not require any viral intermediates or host genome integration during reprogramming. Advantages of this strategy:
The novel iPSC reprogramming strategy we developed based on episomal vectors allows the derivation of transgenic-free iPSCs. Moreover, we have developed new minicircle vectors to express reprogramming factors as non-integrating and non-replicating episomes. Advantages of this strategy include:
The field of cell reprogramming is rapidly expanding. Safe and effective reprogramming methods are becoming increasingly important for both basic research and medical applications. Creative Bioarray provides a wide range of cell reprogramming services to our customers while also helping them develop efficient gene delivery and expression systems for the efficient and rapid generation of iPSCs. If you need our scientific services, please contact us directly.
Reference
- Nishimura, K.; et al. Simple and effective generation of transgene-free induced pluripotent stem cells using an auto-erasable Sendai virus vector responding to microRNA-302. Stem cell research. 2017, 23: 13-19.