Part:BBa_K3335000

We use KIBRA to control exosome secretion

KIBRA as an adaptor-like protein that stabilizes Rab27a, which in turn controls exosome secretion. Rab27a is stabilized by interacting with KIBRA, which prevents ubiquitination and degradation via the ubiquitin-proteasome pathway. KIBRA controls exosome secretion via inhibiting the proteasomal degradation of Rab27a[1].

In our experiment, we used KIBRA to promote cell release of exosomes containing siRNA, thus increasing the efficiency of cell production and release of exosomes and improving the killing of tumor cells.

[1]Lin Song et al,.KIBRA controls exosome secretion via inhibiting the proteasomal degradation of Rab27a.Nature Communication,2019.

Contribution: NJU-China 2020

As previously reported, CMV-hSDC4-STEAP3-NadB can strongly promote exosome release. To develop the more efficient booster part for our strategy, we designed another two parts (CMV-KIBRA and CMV-nSMase2) to find out the most efficient design for our project. We tested all the three designs in vitro. The expression of each mRNA was confirmed by RT-qPCR after transfected to HEK293T cells.

Figure 1. KIBRA, NadB, nSMase2 mRNA relative expression in HEK293T cell (vs GADPH)

KIBRA protein overexpressed in HEK293T

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 335
  Illegal BglII site found at 2999
  Illegal BamHI site found at 246
  Illegal BamHI site found at 442
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 435
  Illegal BsaI site found at 2598
  Illegal BsaI.rc site found at 1920
  Illegal BsaI.rc site found at 2644