Akademik Veri Yönetim Sistemi
11th International Molecular Biology and Biotechnology Congress, Adana, Türkiye, 10 Ekim 2022, cilt.1, ss.56
Spinal muscular atrophy (SMA), a genetically based neuromuscular disease, is an autosomal recessive disease
progressed by the loss of motor neuron cells. The incidence of SMA, which is classified as an inherited monogenic
disease, is higher than other rare diseases. SMA is mostly caused by mutations in the SMN1 gene. With the deletion
of SMN1, neuronal cell loss occurs. Although there is currently no effective treatment for SMA , some degree of
success has been achieved in curing the disease by increasing the SMN2 protein level or by stimulating the
production of SMN1 proteins in the cell. Since a particular gene or gene groups are affected in genetic diseases, it
is important to use a targeted modification method to develop a potential treatment. Mutations in SMN1 could be
reversed by genome editing technologies. Today, CRISPR-Cas systems are widely used because of both their high
targeting and their advantages over some other methods used in genome editing. The use of this approach on
diseases such as SMA will create an opportunity for treatment. However, presenting a heterogeneous mixture as a
package creates the greatest difficulty. The use of exosomes, which are secreted by the cell and have an important
role in intercellular communication, is focused in this study to demonstrate the usability of exosomes as the carrier
of this large heterogeneous complex. It is aimed to carry the CRISPR-Cas9 system and healthy SMN1 sequence
to the cells via exosomes and to make necessary modifications in the cells by using exosomes in rat adipose tissuederived
mesenchymal stem cells (AT-MSC) carrying the experimentally created Smn1-/- phenotype. After CD90+,
CD73+, CD105+, CD34-, HLA-DR-, CD45- AT-MSC were obtained and their differentiation capacities were
characterized, CD9+ and CD81+ exosomes were obtained from these cells. Using cationic polymers, exosome
contents were improved by delivering CRISPR-Cas9-SMN1 vector and healthy SMN1 exon sequence. After the
colon purification, the purified exosome structures were transferred to AT-MSC culture to remove the SMN1
deletion mutation sequence and replace it with a healthy SMN1 sequence (homologous recombination). Fusion of
exosomes into cells was observed by labeling, and it was observed that the SMN1 gene deletion mutation was
eliminated and the healthy SMN1 gene was inserted into the genome, shown by sequence analysis. In this study,
gene editing vector system and healthy SMN1 were transferred by using exosomes as transport vehicles. It has
been shown that cells carrying the healthy SMN1 gene can be obtained in vitro after the culture. Thus, a possible
gene therapy approach that can be used for SMA has been studied in vitro.
This study was supported by TUBİTAK 1003 Projects Support Program Molecular Medicine Call (Project No.
216S467).