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Project Title2011-078 Edible Transgenic Plants as Oral Delivery Vehicles for RNA-based Therapeutics
Track Code2011-078
Websitehttp://otc.rutgers.edu/
Short Description

Rutgers scientists have developed compositions and methods for the production of edible plants expressing small RNAs (sRNAs), such as small interfering RNAs (siRNAs), which are important for down-regulation of therapeutic targets.

Abstract

Rutgers scientists have developed compositions and methods for the production of edible plants expressing small RNAs (sRNAs), such as small interfering RNAs (siRNAs), which are important for down-regulation of therapeutic targets.
Specifically, Rutgers scientists have constructed several binary vectors using:
a) a 500 base pair (bp) fragment from the conserved
nucleoprotein (NP) gene of the influenza virus
H1N1 (IFV);
b) a 498 bp fragment of the core protein sequence from
the hepatitis C virus (HCV) genome; and
c) a 306 bp fragment of the Tat-encoding sequence of
human immunodeficiency virus (HIV).

The constructs were used to create transgenic tomato lines using the commercial cultivar Moneymaker as the genetic background. Each plasmid construct introduced into tomato plants has a nptII marker gene that confers kanamycin resistance and a bacterial β-glucuronidase (GUS) gene as a reporter gene to help screen the transformed tomato plants. All tomato plants were tested for transgene activity by rapid leaf tissue histochemical staining and PCR genotyping, and four plants from each of the three groups of transgenic plants, designated as HIV interfering (HIVi), IFV interfering (NPi), and HCV interfering (HCVi), with different levels of GUS expression were selected and asexually propagated.

• Studies with purified sRNAs transfected into mammalian cells showed that tomato-expressed siRNAs down-modulate the targeted viral sequences in the cells with a high degree of specificity and efficacy


• Parallel studies with rabbits fed with the transgenic tomato fruits showed that sRNAs in fruits can be efficiently taken up into the circulatory system of mammals after ingestion, making this approach a viable oral delivery method

• Further time course studies after termination of tomato feeding indicated that the plant siRNAs can remain stable in rabbits for more than 2 weeks 

• Additional studies are currently being performed to demonstrate efficacy for viral resistance in small animal models as well as to determine dosage dependence.

 
TagsAgricultural Biotechnology; Pharmaceutical; Anti-viral; Small RNA Delivery for Therapeutics; Foods; Vegetables, etc.
 
Posted DateMay 7, 2012 3:46 PM

Researcher

Name
Eric Lam

Manager

Name
Leon Segal

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