Breast cancer, as the most common cancer in women worldwide, represents about 30% of all cancers affecting women. Long non-coding RNAs (lncRNAs) have been implicated in the regulation of several biological processes, and their dysregulation in cancer has well been documented. To investigate possible age-dependent variations in expression profiles of lncRNAs, we evaluated the expression levels of four lncRNAs, i.e., MALAT1, GAS5, SRA, and NEAT1, in breast cancer (BC) samples obtained from younger (<45 years) and older (>45 years) women. Tumor samples (n = 23) and 15 normal tissues were collected from BC patients. All tumor and normal samples were morphologically confirmed by a pathologist. RNA was extracted from the tissues and cDNAs were then synthesized. The lncRNA expression levels were evaluated by qRT-PCR. The changes in the expression levels were determined using the ΔΔCt method. Compared to normal tissues, BC tissues from both age groups (women under 45 years of age and women above 45 years of age) showed upregulation of MALAT1 (p = 0.003 and p = 0.0002), SRA (p = 0.005 and p = 0.0002), and NEAT1 (p = 0.010 and p = 0.0002) and downregulation of GAS5 (p = 0.0002 and p = 0.0005). Additionally, our analysis showed significant and direct correlation between the age and the expression levels of three of the four lncRNAs studied in this work. All four lncRNAs were overexpressed in both MDA-MB-231 and MCF7 cell lines (p = 0.1000). Our data show that MALAT1, GAS5, SRA, and NEAT1 lncRNAs are dysregulated in BC samples. However, except for MALAT1, the expression levels of all of these lncRNAs were significantly lower in cancers developed in younger cases, where poorer prognosis is suggested. Of note, GAS5 reduced expression has been documented to correlate with tumor progression.

Integration target site is the most important factor in successful production of transgenic animals. However, stable expression of transgene without disturbing the function of the host genome depends on promoter methylation, transgene copy number and transcriptional activity in integration regions. Recently, new genome-editing tools have made much progress, however little attention has been paid to the identification of genomic safe harbors. The aim of the present study was to evaluate the effect of insertion site, promoter and copy number of transgene on the production of embryos from cattle fibroblast cells following somatic cell nuclear transfer (SCNT). So, three donor vectors were constructed with EGFP gene under control of different promoters. Each vector was integrated into safe and nonsafe harbors in the genome using phiC31 integrase. Transgenic clones with a single copy of each vector were isolated. Each clone was analyzed to find site and frequency of integration, expression level and promoter methylation before SCNT, as well as transgene expression level and blastocyst formation rate after SCNT. The data obtained demonstrated that BF5, as a safe harbor, not only showed a stable expression, but also the rate of in vitro-produced embryos from BF5-clones are similar to that of nontransfected cells.Transgenic farm animals such as goats, sheep, and cows are important biomaterials for biomedical and life science researches, including basic research, protein production, and animal models for human diseases 1,2 . The current approaches to the generation of transgenic animals are often inefficient with a low integration rate and variable expression levels due to random and unstable integration of transgene into chromosomal DNA, position effects and the number of inserted copies 1,3 . In order to overcome these defects, the somatic cell nuclear transfer (SCNT) technology has been developed in combination with the site-specific integration of foreign DNA 4 . SCNT technology can effectively increase the efficiency of the production of transgenic farm animals, but cannot overcome the concern about the integration of foreign DNA, which can be resolved by means of phiC31 integration system 2,3 . PhiC31 is a site-specific integrase derived from actinophage QC31 of Streptomyces and employed as a powerful genetic tool for efficient non-viral delivery of transgene to the host chromosomal DNA 5,6 . The integrase is a member of DNA recombinase family that encoded within the genome of Streptomyces bacteriophage. This enzyme is functional in mammalian cells thus it can mediate recombination reaction between the attP or pseudo-attP site in the host genome and attB site in the plasmid. This serine integrase can specifically integrate the DNA containing attB site into attP and produce DNA element flanked by attL and attR sequences which are no longer identified by phiC31 integrase 2,7 . This system has several useful properties, including unidirectional site-specific integration, stable gene expression, integration into the saf...