Molecular imaging of gene expression in living subjects by spliceosome-mediated RNA trans-splicing

doi:10.1073/pnas.0402772101

Molecular imaging of gene expression in living subjects by spliceosome-mediated RNA trans-splicing

^*Department of Radiology and Bio-X Program, Stanford University, James H. Clark Center E150, 318 Campus Drive, Stanford, CA 94305-5427; and ^†Intronn Inc., 910 Clopper Road, South Building, Suite 210, Gaithersburg, MD 20878

Communicated by Michael E. Phelps, University of California School of Medicine, Los Angeles, CA, April 19, 2004 (received for review February 2, 2004)

Abstract

Spliceosome-mediated RNA trans-splicing (SMaRT) provides an effective means to reprogram mRNAs and the proteins they encode. SMaRT technology has a broad range of applications, including RNA repair and molecular imaging, each governed by the nature of the sequences delivered by the pre-trans-splicing molecule. Here, we show the ability of SMaRT to optically image the expression of an exogenous gene at the level of pre-mRNA splicing in cells and living animals. Because of the modular design of pre-trans-splicing molecules, there is great potential to employ SMaRT to image the expression of any arbitrary gene of interest in living subjects. In this report, we describe a model system that demonstrates the feasibility of imaging gene expression by transsplicing in small animals. This represents a previously undescribed approach to molecular imaging of mRNA levels in living subjects.

Footnotes

↵ ‡ To whom correspondence should be addressed. E-mail: sgambhir{at}stanford.edu.
Abbreviations: SMaRT, spliceosome-mediated RNA trans-splicing; PTM, pre-trans-splicing molecules; RASON, radiolabeled antisense oligonucleotide; HPV, human papillomavirus; N2a, neuro-2a; ROI, region of interest; sr, steridian.