CONSTRUCTION OF RECOMBINANT LENTIVIRAL VECTOR CONTAINING SHRNA AGAINST MOUSE FNDC5 GENE IN ORDER TO TRANSDUCE MOUSE EMBRYONIC STEM CELLS

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NAZEM SHIMA; GHAEDI KAMRAN; BABASHAH SADEGH; SADEGHIZADEH MAJID; NASR ESFAHANI MOHAMMAD HOSSEIN

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Journal: Pathobiology Research Year:2016 | Volume:19 | Issue:2 Page(s): 59-73

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Title

CONSTRUCTION OF RECOMBINANT LENTIVIRAL VECTOR CONTAINING SHRNA AGAINST MOUSE FNDC5 GENE IN ORDER TO TRANSDUCE MOUSE EMBRYONIC STEM CELLS

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Keywords

FNDC5Q1

SHRNAQ1

LENTIVIRAL VECTORSQ1

RNAIQ1

Abstract

Objective: RNA interference (RNAi) is considered a potential approach to knock down target genes and their functional assessment. LENTIVIRAL VECTORS serve as an efficient tool to transduce foreign genes in a wide variety of mammalian cells. Fibronectin type I domain-containing 5 (FNDC5) is a glycosylated membrane protein whose transcript levels increase during neural and cardiac differentiation of mouse embryonic stem cells (mESCs). Here, we report the efficacy of FNDC5 gene silencing in mESCs using LENTIVIRAL VECTORS that express SHRNA.Methods: Two distinct SHRNA sequences that targeted FNDC5 coding sequence (CDS) and one scramble SHRNA sequence as a negative control were designed and commercially synthesized. Synthetic SHRNA oligonucleotides were cloned into a lentiviral inducible vector after annealing downstream of the tetracycline inducible H1 promoter. The recombinant lentiviral vector was packaged in HEK293T cells, then mESCs were transduced by lentiviral particles. Expression of SHRNA in transduced cell lines was induced by doxycycline treatment for 48 h.Results: Evaluation of transcript levels of FNDC5 by real-time PCR showed a significant decrease in transduced cells by a mixture of two SHRNAs.Conclusion: Taken together, lentiviral-mediated RNAI that targeted the FNDC5 gene could be considered an efficient tool to silence gene expression in the transduced cell line to study the function of FNDC5.

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APA: Copy

NAZEM, SHIMA, GHAEDI, KAMRAN, BABASHAH, SADEGH, SADEGHIZADEH, MAJID, & NASR ESFAHANI, MOHAMMAD HOSSEIN. (2016). CONSTRUCTION OF RECOMBINANT LENTIVIRAL VECTOR CONTAINING SHRNA AGAINST MOUSE FNDC5 GENE IN ORDER TO TRANSDUCE MOUSE EMBRYONIC STEM CELLS. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), 19(2), 59-73. SID. https://sid.ir/paper/81032/en

Vancouver: Copy

NAZEM SHIMA, GHAEDI KAMRAN, BABASHAH SADEGH, SADEGHIZADEH MAJID, NASR ESFAHANI MOHAMMAD HOSSEIN. CONSTRUCTION OF RECOMBINANT LENTIVIRAL VECTOR CONTAINING SHRNA AGAINST MOUSE FNDC5 GENE IN ORDER TO TRANSDUCE MOUSE EMBRYONIC STEM CELLS. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES)[Internet]. 2016;19(2):59-73. Available from: https://sid.ir/paper/81032/en

IEEE: Copy

SHIMA NAZEM, KAMRAN GHAEDI, SADEGH BABASHAH, MAJID SADEGHIZADEH, and MOHAMMAD HOSSEIN NASR ESFAHANI, “CONSTRUCTION OF RECOMBINANT LENTIVIRAL VECTOR CONTAINING SHRNA AGAINST MOUSE FNDC5 GENE IN ORDER TO TRANSDUCE MOUSE EMBRYONIC STEM CELLS,” PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), vol. 19, no. 2, pp. 59–73, 2016, [Online]. Available: https://sid.ir/paper/81032/en

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