| dc.description.abstract | The immortalization of a cell is a process in which a cell is transformed from its primary
state to the immortal one. Likewise, numerous immortalized cell lines of many living
organisms have been produced to maintain the cell’s characteristics for long-term
projects. Additionally, understanding the genetic mechanism related to immortalization
can provide further insights to improve the quality of immortal cells. However, the
genetic basis of immortalizing process in pigs has not been completely clarified yet as
well as its transcriptome profiles. Nevertheless, RNA sequencing has made it possible
to widen our horizons of the transcriptome by studying gene expression, confirming
pathogenicity and detecting transcript alterations. Hence, in this project, we performed
the variant identification of two primary alveolar macrophages (PAM) compared with
two immortalized porcine alveolar macrophages (iPAM) which were transduced with
two exogenous genes (SV40LT and hTERT) and predicted its consequences involving
immortalizing process. RNA sequencing was used for single nucleotide
polymorphisms (SNPs) and short insertions and deletions (INDELs) identifying in 2
porcine cell lines (immortal; iPAM61 and iPAM303 versus mortal; PAM61 and
PAM303). Using an in-house pipeline, a total of 20,194 known RNA-seq variants were
genotyped, of which a variety of variants were exclusively found in iPAMs. In addition,
functional analysis was carried out using DAVID bioinformatic tool. Interesting
missense gene variants and mutations located in coding regions were explored in many
genes related to mismatch repair such as: LIG1; MSH2; RFC2; PMS2. All in all, we did
not identify any gene mutations in coding regions related to the development of
immortalization, we predicted that non-coding areas might have effect on that process. | en_US |
