TY - JOUR ID - 7070 TI - Expression of Recombinant Alpha-1 Antitrypsin in CHO and COS-7 Cell Lines Using Lentiviral Vector JO - Iranian Journal of Biotechnology JA - IJB LA - en SN - 1728-3043 AU - Ghaedi, Mahboobe AU - Sahebghadam Lotfi, Abbas AU - Soleimani, Masoud AU - Shamsara, Mehdi AU - Arjmand, Sare AU - Adibi, Behzad AD - Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, I.R. Iran AD - Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, I.R. Iran AD - Department of Bacteriology and Virology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran AD - School of Pharmacy, Zanjan University of Medical Sciences, P.O. Box 45195-1338, Zanjan, I.R. Iran Y1 - 2009 PY - 2009 VL - 7 IS - 3 SP - 148 EP - 156 KW - Alpha-1 antitrypsin KW - Transfection KW - Protein production KW - Lentiviral vector DO - N2 - In this study, in order to facilitate and accelerate the production of eukaryotic protein alpha 1-antitrypsin (AAT) with correct post-translational modifications, a protein production system based on the transduction of CHO and COS-7 cells using lentiviral vectors was developed. Human AAT cDNA was cloned into a replication-defective lentiviral vector. The transgene AAT-Jred chimer was transferred to CHO and COS-7 cell lines using this vector and its expressions were visualized by fluorescent microscopy. The mRNA expression levels of the AAT genes were determined using Revearse Transcriptase-Polymerase Chain Reaction (RT-PCR) and its secretion into the medium by both cell types was determined using ELISA. The results show that by employing a lentiviral vector, efficient genetic loading of CHO and COS-7 cells with the AAT gene was achieved. In conclusion, by using a Lentivirus-based gene delivery system, large amounts of recombinant human AAT protein were expressed in both CHO and COS-7 cell lines. This expression system possesses key properties that ensure its application in the delivery of therapeutic genes into mammalian cultured cells. UR - https://www.ijbiotech.com/article_7070.html L1 - https://www.ijbiotech.com/article_7070_3265ba4e828b65e6928d9148e96fe25b.pdf ER -