Plasmid constructs encoding high performance reprogramming factors for the generation of iPS cells
Description:
Each construct contains a single or multiple engineered reprogramming factors (fusions between VP16 activator domain and Oct4, Sox2 and Nanog) in retroviral, inducible lentiviral and episomal vectors respectively. With these engineered reprogramming factors, iPS cells can be generated from both mouse and human somatic cells with enhanced efficiency.
Features:
Full-length cDNAs of murine and human Oct4, Sox2 and Nanog genes were ligated with cDNA encoding the VP16 activation domain (amino acids 446-490; from MLGDG to DEYGG) with a region encoding a glycine-rich linker (TSGLGGGSGGGGSGGGGSG, for Oct4 and Sox2) or without the linker (Nanog). Fusion genes were cloned into retroviral vector pMXs (Takahashi, K. & Yamanaka, S. Cell 126, 663-76, 2006) and inducible lentiviral vector pLV-TRE-EF1a-GFP (Wu, Z. et al.. J Mol Cell Biol 1, 46-54, 2009). pLV-TRE-EF1a-GFP is a lentiviral vector for doxycycline (DOX)-inducible transgene expression with EGFP driven by the EF-1 alpha promoter. The expression of reverse tetracycline transactivator (rtTA) was under the control of EF-1 alpha promoter in a separate lentiviral construct.
For the construction of episomal vector pCEP4-XKYZ-2A, coding sequences for OCT4-VP16 (X), KLF4, SOX2-VP16 (Y) and NANOG-VP16 (Z) were linked in this order with 2A elements (Okita, K., Nakagawa, M., Hyenjong, H., Ichisaka, T. & Yamanaka, S. Science 322, 949-53, 2008) and subcloned into pCEP4 (Invitrogen).
Reference:
Wang Y. et al., Reprogramming of mouse and human somatic cells by high-performance engineered factors. EMBO reports 12, 373-378.
This product is for research use. No claim or representation is intended for its use to provide information for the diagnosis, prevention, or treatment of a disease
Order information:
Part # |
Product Name |
Description |
Quantity |
Price |
P 11031 |
pMXs-X |
pMXs-Oct4 (mouse)-VP16-Flag |
3 ¦Ìg |
Inquire |
P11032 |
pMXs-X3 |
pMXs-Flag-Oct4 (mouse)-3¡ÁVP16 |
|
Inquire |
P 11033 |
pMXs-Y |
pMXs-Sox2 (mouse)-VP16-Flag |
|
Inquire |
P 11034 |
pMXs-Z |
pMXs-Nanog (mouse)-VP16-Flag |
|
|
P 11035 |
pMXs-K |
pMXs-Klf4 (mouse) |
|
|
P 11036 |
TRE-X |
Lenti-EF1-EGFP-tetO-OCT4 (human)-VP16-Flag |
|
|
P 11037 |
TRE-Y |
Lenti-EF1-EGFP-tetO-SOX2 (human)-VP16-Flag |
|
|
P 11038 |
TRE-Z |
Lenti-EF1-EGFP-tetO-NANOG (human)-VP16-Flag |
|
|
P 11039 |
TRE-K |
Lenti-EF1-EGFP-tetO-KLF4 (human) |
|
|
P 11040 |
rtTA |
Lenti-EF1-rtTA-IRES-EGFP |
|
|
P 11041 |
pCEP4-XKYZ-2A |
pCEP4-X (OCT4 (human)-VP16-Flag)-2A-KLF4 (human)-2A-Y (SOX2 (human)-VP16-Flag)-2A-Z (NANOG (human)-VP16-Flag) |
|
|
|