In vitro and in vivo gene silencing by TransKingdom RNAi (tkRNAi).

Methods Mol Biol

Skip Ackerman Center for Molecular Therapeutics, Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.

Published: April 2009

AI Article Synopsis

  • RNA interference (RNAi) is a natural and highly conserved process that can specifically silence genes, showing potential for treating various diseases, but faces challenges like delivery and cost of short interfering RNAs (siRNA).
  • Researchers have developed a method using nonpathogenic bacteria to activate RNAi in mammalian cells, known as TransKingdom RNA interference (tkRNAi), which simplifies delivery and reduces synthesis costs.
  • This tkRNAi approach not only allows for long-term gene silencing and high-throughput genomic studies but also offers a safe and effective way to use RNAi in clinical settings.

Article Abstract

RNA interference (RNAi) is a potent and specific mechanism for eliminating the mRNA of specific genes. This gene silencing mechanism occurs naturally and is highly conserved from plants to human cells, holding promise for functional genomics and for revolutionizing medicine due to its unlimited potential to treat genetic, epigenetic, and infectious disease. However, efforts to unleash the enormous potential of RNAi have met with significant challenges. Delivery is problematic because short interfering RNAs (siRNA) are negatively charged polymers that inefficiently enter cells and undergo rapid enzymatic degradation in vivo. In addition, the synthesis of siRNAs is expensive for long-term research and therapeutic applications. Recently, we have shown that nonpathogenic bacteria can be engineered to activate RNAi in mammalian cells (TransKingdom RNA interference; tkRNAi). This new approach offers several advantages and has significant implications. First, this method allows the establishment of a long-term stable gene silencing system in the laboratory against genes of interests in vitro and in vivo, and enables high-throughput functional genomics screening in mammalian systems. RNAi libraries can be constructed, stored, reproduced, amplified, and used with the help of E. coli as currently done with gene cloning. Second, this technology provides a clinically compatible way to achieve RNAi for therapeutic applications due to the proven clinical safety ofnonpathogenic bacteria as a gene carrier, tkRNAi also eliminates the siRNA manufacture issue, and may circumvent or mitigate host interferon-like responses since siRNA is produced intracellularly.

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http://dx.doi.org/10.1007/978-1-60327-547-7_7DOI Listing

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