MIPS Molecular Imaging Program at Stanford Jianghong Rao

MIPS Molecular Imaging Program at Stanford Jianghong Rao Lucas P 097 jrao@stanford. edu 6 -8563 Stanford University School of Medicine Department of Radiology

A unified reporter gene In cell culture In Vivo In vitro Alkaline Phosphatase b-Gal, Luciferase GFPs MIPS Molecular Imaging Program at Stanford GFPs HSV 1 -t. K Transferrin Luciferase Stanford University School of Medicine Department of Radiology

b-Lactamase as the reporter Small (29 k. D) and monomeric Non-toxic in mammalian cells No orthologue in mammalian cells Large signal amplification MIPS Molecular Imaging Program at Stanford University School of Medicine Department of Radiology

b-Lactamase probes for multimodality imaging In vivo whole animals 1. In vitro setting PET, Near Infrared Fluorescence Ex vivo MIPS Molecular Imaging Program at Stanford University School of Medicine Department of Radiology

Bla (-) Bla (+) Pre-injection 2 h MIPS Molecular Imaging Program at Stanford 4 h 12 h 24 h 48 h 72 h Stanford University School of Medicine Department of Radiology

Quantum Dots a simple linear relationship between crystal size and color: the smaller the size, the shorter the wavelength. MIPS Molecular Imaging Program at Stanford University School of Medicine Department of Radiology

In vitro multiplexing imaging of QD-Luc 8 conjugates Fluorescence MIPS Molecular Imaging Program at Stanford Bioluminescence (BRET) Stanford University School of Medicine Department of Radiology

Multiplexing imaging of QD-Luc 8 conjugates a QD 800 _luc 8 QD 705 _luc 8 QDmix _luc 8 QD 655 _luc 8 open filter b c Ds. Red filter X-cy 5. 5 filter MIPS Molecular Imaging Program at Stanford e d ICG filter fluorescence Stanford University School of Medicine Department of Radiology

Ribozyme for RNA-based gene therapy Group I intron ribozyme from Tetrahymena • Catalytic RNA molecule • Can correct defective gene transcripts • A promising agent for gene therapy MIPS Molecular Imaging Program at Stanford University School of Medicine Department of Radiology

Trans-splicing ribozymes for gene therapy N X Correct transcript Defective gene transcript (mutation site X) NNNNNG N X NNNNNG Ribozyme-catalyzed cleavage & ligation N Corrected transcript MIPS Molecular Imaging Program at Stanford University School of Medicine Department of Radiology

Imaging p 53 expression CCF 2/AM Ds. Red CMV-bla MIPS Molecular Imaging Program at Stanford p 53 mt+ TRz 200 dead Stanford University School of Medicine Department of Radiology