Supplemental materials for: Broxmeyer et al
Files in this Data Supplement:
- Figure S1. Self-inactivating lentiviral vectors were cloned by inserting a fragment consisting of two transcription factors with 2A sequence, internal ribosome entry site (IRES), and fluorescent protein into the NheI and XhoI sites of pCSC-SP-PW-GFP.1 (JPG, 289 KB) -
Expression is driven by the CMV early enhancer/chicken beta-actin promoter (CAGp). The 2A sequence permits the transcription factors to be translated into two separate proteins.2 The 5′ transcription factors will have amino acids --GATNFSLLKLAGDVELNPG on the C-terminus and the 3′ transcription factors will have a proline on the N-terminus after 2A processing. Different fluorescent proteins allows for dual detection in transduced cells. LTR, long terminal repeat; Ψ, packaging signal; RRE, rev response element, WPRE, woodchuck hepatitis post-transcriptional regulatory element.
REFERENCES
1. Marr RA, Guan H, Rockenstein E, et al. Neprilysin regulates amyloid Beta peptide levels. J.Mol.Neurosci. 2004;22:(1�2)5�11.
2. Donnelly ML, Luke G, Mehrotra A, et al. Analysis of the aphthovirus 2A/2B polyprotein �cleavage� mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal �skip.� J.Gen.Virol. 2001;82:(Pt 5)1013�1025.