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June 12-13, 2019 | Edinburgh, Scotland
8th European Clinical Microbiology and Immunology Congress
&
3rd World congress on Biotechnology
Joint Event
Microbiology: Current Research | Volume: 3 | ISSN: 2591-8036
Design and construction of a genetic vector expressing a poly-miR-122 for gene
therapy of hepatocarcinoma
Mariela Montaño-Samaniego
Escuela Nacional de Ciencias Biológicas, México
H epatocellular carcinoma (HCC) is the third leading
cause of cancer-related death worldwide and there is
still no effective treatment for this disease, so gene therapy
is a promising therapeutic alternative for the treatment
of HCC. The use of microRNA (miRNA) in gene therapy
has become a powerful tool for the regulation of genes
involved in acquired genetic diseases such as cancer. The
miRNA-122 (miR-122) is specific and the most abundant
in the liver, it has been shown to function as a tumor
suppressor. The levels of miR-122 decrease significantly
and specifically in HCC. The objective of this work was to
construct a genetic vector that contains a poly-miR-122
governed by the α-fetoprotein (AFP) promoter, specific to
HCC. A poly-miR-122 sequence containing three miR-122
precursors (pre-miR-122) was designed, this was analyzed
for secondary structure prediction and thermodynamic
stability. The result of the prediction analysis of the poly-
miR-122 sequence showed that the primary transcript
will have thermodynamic stability, indicating that it can
function in the treatment against HCC cells. Subsequently,
the recombinant plasmid pIRES2-AFP-poly-miR-122-
EGFP was constructed. The identity of this recombinant
plasmid was confirmed by enzymatic restriction. The
presence of the AFP promoter in the recombinant plasmid
was confirmed by PCR. With automated sequencing, an
identity of 99.6% was found with the AFP promoter (NCBI:
L34019.1). This genetic construction can express the active
and stable miR-122, so it could be used for the specific
treatment of HCC by gene therapy.
e:
Microbiol Curr Res, Volume 3
ISSN: 2591-8036
Notes: