Reprogramming Lentiviruses for Mouse Cells

Stemgent Dox Lentivirus Set: mOKSM (Product code ST000021)

The Stemgent Dox Lentivirus Set: mOKSM provides an affordable inducible method for generating iPS cells. The set includes mOct4, mSox2, mc-Myc, and mKlf4 transcription factors along with reverse tetracycline transcriptional activator (rtTA) and GFP, prepackaged as lentivirus particles tested and proven to reprogram mouse somatic cells. Each unique virus is provided as a VSV-G pseudotyped virus stock capable of infecting both dividing and non-dividing cells. The expression of the four transcription factors, in conjunction with rtTA, has been shown to direct a variety of differentiated cell types to become reprogrammed iPS cells¹.

Stemgent Dox Lentivirus Set: mOKSM (Product code ST000021)

The Stemgent Dox Lentivirus Set: mOKSM provides an affordable inducible method for generating iPS cells. The set includes mOct4, mSox2, mc-Myc, and mKlf4 transcription factors along with reverse tetracycline transcriptional activator (rtTA) and GFP, prepackaged as lentivirus particles tested and proven to reprogram mouse somatic cells. Each unique virus is provided as a VSV-G pseudotyped virus stock capable of infecting both dividing and non-dividing cells. The expression of the four transcription factors, in conjunction with rtTA, has been shown to direct a variety of differentiated cell types to become reprogrammed iPS cells¹.

Stemgent Dox Lentivirus Set: mOKSM, concentrated (Product code ST000014)

Each doxycycline-inducible lentivirus in this set is capable of expressing one of four mouseb transcription factors (Oct4, Sox2, Klf4 or c-Myc) under the control of the doxycycline (DOX)- inducible tetO operator when transduced into mammalian cells. Each unique virus is provided as a VSV-G pseudotyped and concentrated virus stock capable of infecting both dividing and non-dividing cells. The expression of these four transcription factors, along with the reverse tetracycline transcriptional activator (rtTA), has been shown to reprogram a variety of mouse cell types including fibroblasts, B cells, T cells, and neural precursor cells to an embryonic stem (ES) cell-like state known as the induced pluripotent stem (iPS) cell1. Proper isolation and subsequent injection of these iPS cells into blastocysts can generate mice that carry all four inducible viruses, which can be reactivated upon the addition of DOX2.
The DOX Inducible GFP-Lentivirus is included as a transduction control. The Stemgent Dox Lentivirus Set: mOKSM, concentrated, was developed in the lab of Rudolf Jaenisch, M.D., at the Whitehead Institute at MIT¹. Dr. Jaenisch is a recognized leader in the study of epigenetic regulation of gene expression with numerous publications focused on ES and iPS cellular mechanisms and techniques.

Stemgent Dox Lentivirus Set: m4F2A (Product Code ST000043)

The Stemgent Dox Lentivirus Set: m4F2A expresses each of the four mouse transcription factors (Oct4, Sox2, Klf4 or c-Myc) contained within a single expression cassette under the control of the doxycycline (Dox)-inducible tetO operator. The inclusion of three unique 2A peptide (P2A, T2A, and E2A) sequences (one between each transcription factor) results in “ribosomal skipping”, which allows for the equivalent expression of all four transcription factors from a single vector³. Introduction the four factors from a single virus minimizes the number of proviral integrations required for successful reprogramming. This reduces the risk of insertional mutagenesis eff ects in the resultant induced pluripotent stem (iPS) cells when compared to those generated using four independent viruses. The 4F2A virus is capable of generating iPS cells from both embryonic and adult murine somatic cells, as well as neonatal human foreskin keratinocytes (NHFK)⁴. The virus is provided as a VSV-G pseudotyped and concentrated virus stock capable of infecting both dividing and non-dividing cells. The individual expression of these four transcription factors, along with the reverse tetracycline transcriptional activator (rtTA), has been shown to reprogram a variety of mouse cell types including fi broblasts, B cells, T cells, as well as neural precursors to the embryonic stem (ES) cell-like state known as the iPS cell state. Proper isolation and subsequent injection of these iPS cells into blastocysts has been shown to generate mice that carry all four inducible viruses, which can be reactivated upon the addition of doxycycline.

The Stemgent Dox Lentivirus Set: m4F2A was developed in the lab of Rudolf Jaenisch, M.D., at the Whitehead Institute at MIT3. Dr. Jaenisch is a recognized leader in the study of epigenetic regulation of gene expression with numerous publications focused on ES and iPS cellular mechanisms and techniques.