A double built-in containment strategy for production of recombinant proteins in transgenic rice

PLoS One. 2014 Dec 22;9(12):e115459. doi: 10.1371/journal.pone.0115459. eCollection 2014.

Abstract

Using transgenic rice as a bioreactor for mass production of pharmaceutical proteins could potentially reduce the cost of production significantly. However, a major concern over the bioreactor transgenic rice is the risk of its unintended spreading into environment and into food or feed supplies. Here we report a mitigating method to prevent unwanted transgenic rice spreading by a double built-in containment strategy, which sets a selectively termination method and a visual tag technology in the T-DNA for transformation. We created transgenic rice with an inserted T-DNA that harbors a human proinsulin gene fused with the far-red fluorescent protein gene mKate_S158A, an RNAi cassette suppressing the expression of the rice bentazon detoxification enzyme CYP81A6, and an EPSPS gene as the selection marker for transformation. Herbicide spray tests indicated that such transgenic rice plants can be killed selectively by a spray of bentazon at regular field application dosage for rice weed control. Moreover, the transgenic rice seeds were bright red in color due to the fused far-red fluorescent protein, and could be easily visualized under daylight by naked eyes. Thus, the transgenic rice plants reported in this study could be selectively killed by a commonly used herbicide during their growth stage, and their seeds may be detected visually during processing and consumption after harvest. This double built-in containment strategy may greatly enhance the confinement of the transgenic rice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzothiadiazines / pharmacology*
  • Containment of Biohazards*
  • DNA, Bacterial / genetics
  • Herbicides / pharmacology
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Oryza / genetics*
  • Oryza / growth & development
  • Oryza / metabolism
  • Plant Diseases / microbiology
  • Plant Diseases / prevention & control
  • Plants, Genetically Modified / genetics*
  • Plants, Genetically Modified / growth & development
  • Plants, Genetically Modified / metabolism
  • Proinsulin / genetics
  • Proinsulin / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Seeds / chemistry

Substances

  • Benzothiadiazines
  • DNA, Bacterial
  • Herbicides
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • T-DNA
  • red fluorescent protein
  • Proinsulin
  • bentazone

Grant support

This work was supported by the Grant from the Department of Agriculture of China (2014ZX08011-006). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.