Papers by kianoush dormiani
یازدهمین کنگره ژنتیک ایران, 2010
Scientific Reports, Aug 21, 2023
RNA-binding protein Musashi1 (MSI1) shows an increased expression level in several cancers and ha... more RNA-binding protein Musashi1 (MSI1) shows an increased expression level in several cancers and has been introduced as a prognostic marker in some malignancies. It is expected that if any miRNA is encoded by this gene, it might have a role in cancer development or could be considered as a prognostic biomarker. Accordingly, in this study, we aimed to find novel miRNA(s) inside the intronic regions of the MSI1 gene. Here, we report two novel miRNAs within intron 4 of MSI1 gene, named MSM2 and MSM3, which were selected among several miRNA precursors predicted by bioinformatic studies. For experimental analysis, corresponding precursor miRNAs were transfected into HEK293T cells and exogenous expression of the mature miRNAs were detected. Two mature miRNAs, MSM3-3p and MSM3-5p were generated by MSM3 precursor and one, MSM2-5p was derived from MSM2. Besides, endogenous expression of MSM2-5p and MSM3-3p was detected in MCF-7 and SH-SY5Y cell lines. Expression of both mature miRNAs was also detected in clinical samples of breast cancer. Additionally, the interaction between the MSM3-3p and 3′UTR region of PDE11A was confirmed by dual luciferase assay. Overall, our data demonstrated that MSI1 gene encodes two novel miRNAs in breast cancer cells. miRNAs are small non-coding RNA molecules that bind to the 3′UTR regions of target genes to regulate their expression post-transcriptionally. miRNAs play critical roles in many cellular processes and developmental pathways. It is documented that aberrant expression of miRNAs is associated with several disorders. In cancer biology, miRNAs may play the tumor suppressor or oncogenic roles and their expression profile differs in different stages of a particular cancer 1,2. Also, due to their high stability in different tissues and body fluids, miRNAs are considered as potential non-invasive biomarkers 3,4. Different stages of cancer, including diagnosis/prognosis, remission, relapse and metastasis have specific miRNA signatures, which contain valuable information applicable for the clinical management of cancer. miRNA signature may be important in determining the drug efficacy in cancer treatment 5. For the discovery of novel miRNAs, several approaches including experimental and computation-driven methods are employed. In these two methods, miRNAs with a very low expression rate are not detectable. Moreover, non-conserved miRNAs and some miRNAs due to their complex structure and inability to subclone, are missed 6,7. In order to reduce the possibility of missing novel miRNAs due to the limitations of these methods, in this study we used the combination of both experimental and computation-driven methods to discover novel miRNA(s) inside MSI1 gene. The RNA-binding protein Musashi1 (MSI1) plays a critical role in normal cell proliferation as well as the differentiation and development of several organs. MSI1 protein is highly expressed in stem cells of different tissues and its aberrant expression is reported in many tumors 8,9. MSI1 is considered as an activator in tumorigenesis 10-12 and its increased expression is reported in many cancers such as lung, pancreas, glioma, breast, and colon cancers 13-15. However, the function and mechanism of action of MSI1 are not fully understood. Kawahara et. al. showed that MSI1 interrupts translation initiation of MSI1 target mRNAs by competing with eIF4G for binding to PABPC1 16. MSI1 activates the NOTCH and WNT pathways and consequently increases the cell proliferation and keeps the stemness status of cancer cells 17. Many studies have shown that knockdown of MSI1 reduces cancerous properties including cell proliferation, radioresistance and invasion. MSI1 knockdown in glioblastoma and
Scientia Pharmaceutica, 2021
Introduction: Bioactive encapsulation and drug delivery systems have already found their way to t... more Introduction: Bioactive encapsulation and drug delivery systems have already found their way to the market as efficient therapeutics to combat infections, viral diseases and different types of cancer. The fields of food fortification, nutraceutical supplementation and cosmeceuticals have also been getting the benefit of encapsulation technologies. Aim: Successful formulation of such therapeutic and nutraceutical compounds requires thorough analysis and assessment of certain characteristics including particle number and surface area without the need to employ sophisticated analytical techniques. Solution: Here we present simple mathematical formulas and equations used in the research and development of drug delivery and controlled release systems employed for bioactive encapsulation and targeting the sites of infection and cancer in vitro and in vivo. Systems covered in this entry include lipidic vesicles, polymeric capsules, metallic particles as well as surfactant- and tocopherol-b...
Tenecteplase is a variant of tissue plasminogen activator (t-PA) which has better pharmacokinetic... more Tenecteplase is a variant of tissue plasminogen activator (t-PA) which has better pharmacokinetic properties and more selective thrombolytic activity. In the present study, we describe a rapid method to introduce three sets of mutation into defined positions in t-PA cDNA by a site-directed mutagenesis based on a megaprimer PCR approach to produce tenecteplase coding sequence where amino acids at positions 103, 117 and 296-299 in polypeptide sequence were modified. This sequence was cloned in pTZ57R by T/A cloning method to introduce suitable restriction sites at both ends of the amplified fragment. Vectors containing tenecteplase cDNA were propagated in One Shot TOP10 chemically competent E. coli and positive clones were selected by blue/white screening method. After being isolated and purified, the recombinant plasmids were digested by suitable restriction enzymes. Acquired tenecteplase coding sequence was subcloned into a tissue specific expression vector. Upon expression, it is expected that tenecteplase will be expressed exclusively in lactating mammary glands during milk production in transgenic animal. Finally, the recombinant vector was isolated and verified by restriction digestion and sequence analysis to confirm that the tenecteplase coding sequence has been inserted in the proper orientation.
Cellular Reprogramming, Apr 1, 2011
The purpose of this study was to develop an improved zona-free method of goat somatic cell nuclea... more The purpose of this study was to develop an improved zona-free method of goat somatic cell nuclear transfer (SCNT) that has both ease of operation and efficiency. The main steps involved were: 1) optimization of in vitro oocyte maturation, 2) parthenogenetic activation of zona-free oocytes, 3) SCNT of zona-free anaphase II-telophase II (AII-TII) oocytes which subverted the need for long term UV-exposure of the oocytes, and 4) in vitro culture of groups of cloned embryos in wells in a highly efficient continuous serum-free embryo medium to the blastocyst stage before transfer to the recipients. Percentages of transgenic blastocyst production were 22.3% and 33.1% for adult and fetal cell lines, respectively. After transfer of cloned and transgenic blastocysts, 28.6% and 36.4% of the recipients were confirmed pregnant and 75% and 25% of the pregnancies resulted in the delivery of viable offspring, respectively. To our knowledge this is the first report of successful viable birth of cloned and transgenic offspring through a whole procedure of in vitro oocyte maturation and embryo development to the blastocyst stage, and in this study the in vitro efficiencies of cloned and transgenic embryo production were higher than the available reports.
International Journal of Biological Macromolecules, Mar 1, 2023
Cell journal, 2018
The Streptomyces phage phiC31 integrase offers a sequence-specific method of transgenesis with a ... more The Streptomyces phage phiC31 integrase offers a sequence-specific method of transgenesis with a robust long-term gene expression. PhiC31 has been successfully developed in a variety of tissues and organs for purpose of in vivo gene therapy. The objective of the present experiment was to evaluate PhiC31-based site-specific transgenesis system for production of transgenic bovine embryos by somatic cell nuclear transfer and intracytoplasmic sperm injection. In this experimental study, the application of phiC31 integrase system was evaluated for generating transgenic bovine embryos by somatic cell nuclear transfer (SCNT) and sperm mediated gene transfer (SMGT) approaches. PhiC31 integrase mRNA and protein was produced in vitro and their functionality was confirmed. Seven phiC31 recognizable bovine pseudo attachment sites of phage (attP) sites were considered for evaluation of site specific recombination. The accuracy of these sites was validated in phic31 targeted bovine fibroblasts us...
PLOS ONE
SCNT embryos suffer from poor developmental competence (both in vitro and in vivo) due to various... more SCNT embryos suffer from poor developmental competence (both in vitro and in vivo) due to various defects such as oxidative stress, incomplete epigenetic reprogramming, and flaws in telomere rejuvenation. It is very promising to ameliorate all these defects in SCNT embryos by supplementing the culture medium with a single compound. It has been demonstrated that melatonin, as a multitasking molecule, can improve the development of SCNT embryos, but its function during ovine SCNT embryos is unclear. We observed that supplementation of embryonic culture medium with 10 nM melatonin for 7 days accelerated the rate of blastocyst formation in ovine SCNT embryos. In addition, the quality of blastocysts increased in the melatonin-treated group compared with the SCNT control groups in terms of ICM, TE, total cell number, and mRNA expression of NANOG. Mechanistic studies in this study revealed that the melatonin-treated group had significantly lower ROS level, apoptotic cell ratio, and mRNA ex...
PLOS ONE
SCNT embryos suffer from poor developmental competence (both in vitro and in vivo) due to various... more SCNT embryos suffer from poor developmental competence (both in vitro and in vivo) due to various defects such as oxidative stress, incomplete epigenetic reprogramming, and flaws in telomere rejuvenation. It is very promising to ameliorate all these defects in SCNT embryos by supplementing the culture medium with a single compound. It has been demonstrated that melatonin, as a multitasking molecule, can improve the development of SCNT embryos, but its function during ovine SCNT embryos is unclear. We observed that supplementation of embryonic culture medium with 10 nM melatonin for 7 days accelerated the rate of blastocyst formation in ovine SCNT embryos. In addition, the quality of blastocysts increased in the melatonin-treated group compared with the SCNT control groups in terms of ICM, TE, total cell number, and mRNA expression of NANOG. Mechanistic studies in this study revealed that the melatonin-treated group had significantly lower ROS level, apoptotic cell ratio, and mRNA ex...
Materials Science and Engineering: C, 2021
A novel polyelectrolyte nanocarrier was synthesized via layer-by-layer self-assembly of polycatio... more A novel polyelectrolyte nanocarrier was synthesized via layer-by-layer self-assembly of polycationic and polyanionic chains. The nanocarrier is composed of polyglutamate grafted chitosan core, dextran sulfate as a complexing agent, and polyethyleneimine shell decorated with folic acid. This polyelectrolyte complex has unique physicochemical properties so that the core is considered as an efficient carrier for LTX-315 and melittin peptides, and the shell is suitable for delivery of miR-34a. The spherical nanocarriers with an average size of 123 ± 5 nm and a zeta potential of -36 ± 1 mV demonstrated controlled-release of gene and peptides ensured a synergistic effect in establishing multiple cell death pathways on chemoresistance human breast adenocarcinoma cell line, MDA-MB-231. In vitro cell viability assays also revealed no cytotoxicity for the nanocarriers, and an IC50 of 15 μg/mL and 150 μg/mL for melittin and LTX-315, respectively, after 48 h, whereas co-delivery of melittin with miR-34a increased smart death induction by 54%.
Applied Microbiology and Biotechnology
Additional file 7: Figure S6. Agarose electrophoresis of double digestion products of recombinant... more Additional file 7: Figure S6. Agarose electrophoresis of double digestion products of recombinant plasmids [pBUD/precursor miRNAs expression plasmids] which were digested with SalI and XbaI restriction enzymes. Double digestion with SalI and XbaI resulted in two distinct bands with approximate sizes of 5191 bp for plasmid pBUD and 378 bp for precursor of miR-637, 453 bp for precursor of miR-802 and 427 bp for precursor of miR-125b. M is 1kbp DNA ladder (Thermo Scientific, USA).
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Papers by kianoush dormiani