Production of Transgenic Murine Blastocyst By DMSO-Sperm Mediated Gene Transfer

سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 585

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شناسه ملی سند علمی:

ISERB04_017

تاریخ نمایه سازی: 16 تیر 1397

چکیده مقاله:

Backgrond: One of the methods with high efficiency and simplicity to produce transgenic mice based on transfect spermatozoa with exogenous DNA/DMSO complex which isnamely called DMSO-sperm mediated gene transfer (SMGT). SMGT based on the ability of the sperm cells to bind to foreign molecules DNA and insert it into yourself and then transfer this molecules in the egg during fertilization. Shin and his colleagues developed a high efficient and simple method to obtain transgenic offspring that’s called DMSO-sperm mediated gene transfer. In this study transgenic murine blastocyst by DMSO-SMGT was produced. Methods: Mouse sperm collected from caudae epididymis mice and incubated in 37◦C for 60 min. Mouse sperm cell suspension (107 cells/ml) was supplemented with the DMSO Of final final3%concentration, circle pEGFP-N1 DNA (20 μg/ml) in HTF. The mixture was cooled to 4◦C for 10_15 min. Then 10 microliters from sperm suspension used to do IVf. After 4_5 h of in vitro fertilization (IVF), inseminated oocytes were incubated in KSOM. After 4 days transgenic blastocyst obtained. Results: Transgenic murine blastocyst Produced by IVF of Sperms Transfected with EGFP Transgenic rate approximately 46% obtained. Conclusion: Despite the advantages of the SMGT method, such as convenience, simplicity and low cost, the efficiency of SMGT is only about 3-6%, researchers trying to find ways to increase the efficiency of this method for producing transgenic animals .DMSO can improve efficiency of SMGT method.

کلیدواژه ها:

نویسندگان

Soleiman Kurd

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences

Sara Hosseini

Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences

Vahid Jajarmi

Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences

Fardin Fathi

Cellular and Molecular Research Center, Kurdistan University of Medical Sciences