In vivo targeted delivery of CD40 shRNA to mouse intestinal dendritic cells by oral administration of recombinant Sacchromyces cerevisiae

Abstract

Short hairpin RNA (shRNA)-mediated gene regulation is a commonly used technique for gene manipulation. An efficient and safe delivery system is indispensable when shRNA is delivered into living organisms for gene therapy. Previous studies have proved that DNA and protein can be delivered into dendritic cells (DCs) by non-pathogenic Saccharomyces cerevisiae without being degraded. CD40 is closely related to apoptosis of tumor cells and some immune mechanisms. In this study, we demonstrated that recombinant yeast S. cerevisiae efficiently delivered the shRNA of immune-associated gene (CD40) into mouse intestinal DCs via oral administration. Western blot analysis of isolated intestinal DCs indicated that the inhibition of CD40 gene expression reached up to 56–91%. The secretion of cytokines such as interleukin-2 (IL-2), IL-6, IL-10, IL-12, tumor necrosis factor-α and interferon-γ in intestinal DCs had varying degrees of changes. In conclusion, we found that orally administered recombinant yeast can be used as an efficient shRNA delivery system for intestinal DC-specific gene silencing and immunomodulation in vivo.

Intestinal DCs global gene expression dynamics affected by recombinant baker’s yeasts saccharomyces cerevisiae

“…The complex p35/CDK5 is required for neurite outgrowth and cortical lamination [37] and involved in dendritic spine morphogenesis by mediating the EFNA1-EPHA4 signaling. Much more than this, as described previously, saccharomyces cerevisiae can be an effective oral DNA vaccine vehicle targeting for intestinal cells and provoke strong immune response in different animal species [38][39][40]. Our results further proved the utility of oral yeast-based gene delivery vehicle targeting for intestinal DCs directly, which lays a solid foundation for the development of yeast-based DNA vaccine.…”

Multiplex CRISPR/Cas9-based genome engineering enhanced by Drosha-mediated sgRNA-shRNA structure

“…To validate that the sgRNA-shRNA transcript can be processed to functional sgRNAs, we generated the msgRNA-2 construct containing VEGF.sgRNA, CCR5.sgRNA and an irrelevant shRNA32 (Fig. 2a, Supplementary Fig.1a and Sequence 1) and the corresponding DsRed-eGFP surrogate reporter vectors (Fig.…”

“…4a,), respectively. The CON.shRNA represented the control irrelevant shRNA as we previously used for mouse CD40 mRNA interference32. The LIG4.shRNA represented the shRNA designed against LIG4 gene.…”

Surface-modified yeast cells: A novel eukaryotic carrier for oral application

“…In the last years, we and other groups described the successful usage of intact, recombinant yeast cells for the delivery of both proteins [17][18][19][20][21][22] and functional nucleic acids [23][24][25][26][27][28] …”