The design and optimization of RNA trans-splicing molecules for skin cancer therapy.

[dystrophic epidermolysis bullosa]

Targeting tumor marker genes by RNA trans-splicing is a promising means to induce tumor cell-specific death . Using a screening system we designed RNA trans-splicing molecules (RTM ) specifically binding the pre-m RNA of SLCO 1 B 3 , a marker gene in epidermolysis bullosa associated squamous cell carcinoma (EB-SCC ). Specific trans-splicing , results in the fusion of the endogenous target mRNA of SLCO 1 B 3 and the coding sequence of the suicide gene , provided by the RTM . SLCO 1 B 3 -specific RTMs containing HSV-tk were analyzed regarding their trans-splicing potential in a heterologous context using a SLCO 1 B 3 expressing minigene (SLCO 1 B 3 -MG ). Expression of the chimeric SLCO 1 B 3 -tk was detected by semi-quantitative RT-PCR and Western blot analysis . Cell viability and apoptosis assays confirmed that the RTMs induced suicide gene -mediated apoptosis in SLCO 1 B 3 -MG expressing cells . The lead RTM also showed its potential to facilitate a trans-splicing reaction into the endogenous SLCO 1 B 3 pre-m RNA in EB-SCC cells resulting in tk -mediated apoptosis . We assume that the pre-selection of RTMs by our inducible cell-death system accelerates the design of optimal RTMs capable to induce tumor specific cell death in skin cancer cells .

Diseases
Validation

Diseases presenting "squamous cell carcinoma" symptom

carcinoma of the gallbladder

child syndrome

dystrophic epidermolysis bullosa

epidermolysis bullosa simplex

esophageal adenocarcinoma

esophageal carcinoma

esophageal squamous cell carcinoma

junctional epidermolysis bullosa

kallmann syndrome

kindler syndrome

liposarcoma

monosomy 21

oculocutaneous albinism

oral submucous fibrosis

papillon-lefèvre syndrome

This symptom has already been validated