LoxP 4-in-1 iPS Factor on Lentiviral Vectors for Efficient Reprogramming

Putting 4 iPS factors on one lentiviral vector, separated by 2A peptides, has appeared to be more efficient in generating iPS cells than having all 4 factors on individual viruses, at least in a number of cases. Stem cell-like colonies start to appear in about 2 weeks using Allele Biotech’s 4-in-1 lentivirus. In addition to the concerted effects from Oct3/4, Sox2, c-Myc, Klf4, it is also believed that the coordinated silencing of these factors after reprogramming help forming iPS colonies.

The 4-in-1 lentivirus from Allele Biotech contains loxP sites that can be used to remove the 4 cDNAs if so desired. For convenience, a new product kit is offered starting this week to include lenti-nCre in a kit with the 4-in-1 iPS viral products.

New Product of the Week 04-11-10 to 04-18-10: 4-In-One-Vector: Human OSKM Lentiviral Paticles (Oct3/4, Sox2, Klf4 and c-Myc) and Cre Lentiviral Particle kits, Cat # ABP-SC-LVI4IN1C1 or ABP-SC-LVI4IN1C5

Promotion of the Week 04-11-10 to 04-18-10: Single vial 4-in-1 is offered only this week. This product has been well established and validated, one of the reasons smaller packages are not normally offered. As a matter of fact, every batch of the 4-in-1 iPS lentivirus has been sold out.

Update note: Lentivirus inserts into the host chromosome, and is gradually being replaced by footprint-free reprogramming reagents, the best being Allele Biotech’s enhanced mRNA reprogramming factors that feature a patent-pending fusion gene.

Tags: 4-in-1, c-myc, cre, iPS, iPSCs, Klf4, lentivirus, loxP, Oct3, repgramming, silencing, Sox2

Reprogramming Life

President Obama is expected to lift the ban on federal fund for embryonic stem cell research soon. However, that does not seem to be the hottest topic these days concerning stem cell research. In 2006, Shinya Yamanaka showed that mouse skin cells could be reprogrammed back into something called induced pluripotent stem (iPS) cells by introducing a handful of cDNAs using retroviral vectors. The process was later repeated in human cells and by other groups including those of Thomson and Melton, sometimes with a slightly different set of inducing cDNAs, or with chemicals or shRNA repressing the repressors of the inducer genes.

The iPS cells are not exactly the same as ES cells, and no animals have been created using iPS cells, but they are close enough to be of great interest to lots of people, particularly for basic research purposes. The method to create iPS by reversing chromosomal changes along differentiation pathways appears to be surprisingly simple, like erasing an old audio tape, there may still be acoustic information left if analyzed by the right equipment, but to most people it is as clean as new. You’d wish a few things in life could be reversed that easily!

For labs that are not already in the stem cell field but feel a need to get their feet wet, then they want reagents that are pre-assembled and pre-tested. Such reagents may include: iPS cultures, iPS inducing viral particles, antibodies to stem cell specific markers, cell assays, and even PCR primer sets (synthesizing hundreds of oligos used in the Yamanaka papers alone will take a lot time and unnecessary costs). That’s where a fast-moving, research-oriented company like Allele comes in. We will bring what we think as starter sets for you, and listen to what you think as needed as we along. The new iPS product line will be launched within weeks, hopefully coinciding with our brand new webpages for all our current product lines!

Tags: c-myc, desease tissue regeneration, differetiation, iPS, Klf4, Lin28, Nanog, Oct3/4, pathways, pluripotent, re-programming, Sox2, stem cells, therapy