Optimization of mouse in utero electroporation technique

YANG Xiao-wei LIU An-jun REN An-jing WANG Zhi-BinZHANG Wei-ping Lü Jian-xin* XIE Zhi-fang*

doi:10.16098/j.issn.0529-1356.2018.03.020

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Acta Anatomica Sinica ›› 2018, Vol. 49 ›› Issue (3) : 391-394. DOI: 10.16098/j.issn.0529-1356.2018.03.020

Technology and Methodology

Optimization of mouse in utero electroporation technique

YANG Xiao-wei ^1,2 LIU An-jun ^2,3 REN An-jing² WANG Zhi-Bin²ZHANG Wei-ping² Lü Jian-xin ^1* XIE Zhi-fang ^2*

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Abstract

Objective To optimize the in utero electroporation protocol for transfering genes into the embryonic murine brain. Methods Critical steps of in utero electroporation were optimized, including choosing isoflurane anesthesia, improving the preparation of micropipettes, reducing the exposure time of embryos during the surgery as well as optimizing electroporation parameters. Forty-two mouse embryos from 6 mothers were subjected to in utero electroporation at gestational day 16.5 using the optimized protocol. Results The survival rates of mothers and embryos were 100%. The tranfered gene was expressed in 85%±3% of electroporated embryonic brains. Conclusion Using the optimized in utero electroporation protocol, a satisfying survival rate of embryos and transfection efciency can be achieved.

Key words

Brain / Gene / Development / In utero electroporation / Mouse

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YANG Xiao-wei LIU An-jun REN An-jing WANG Zhi-BinZHANG Wei-ping Lü Jian-xin XIE Zhi-fang. Optimization of mouse in utero electroporation technique[J]. Acta Anatomica Sinica. 2018, 49(3): 391-394 https://doi.org/10.16098/j.issn.0529-1356.2018.03.020

References

［1］Shinmyo Y, Kawasaki H. CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation［J］. Curr Protoc Neurosci, 2017, (79): 3-32.1-3-32.11.

［2］Tsunekawa Y, Terhune RK, Fujita Ⅰ, et al. Developing a de novo targeted knock-in method based on in utero electroporation into the mammalian brain［J］. Development, 2016, 143 (17): 3216-3222.

［3］Saito T. In vivo electroporation in the embryonic mouse central nervous system［J］. Nat Protoc, 2006, 1 (3): 1552-1558.

［4］Tabata H, Nakajima K. Labeling embryonic mouse central nervous system cells by in utero electroporation［J］. Dev Growth Differ, 2008, 50 (6): 507-511.

［5］Yang CQ, Li XY, Lu X, et al. Construction of in utero electroporation in the mouse［J］. Chinese Journal of Anatomy, 2013, 36 (2): 170-172. (in Chinese)

杨慈清，李小英，卢习, 等.小鼠子宫内电转方法的建立［J］. 解剖学杂志，2013, 36 (2):170-172.

［6］Wu Y, Ding XF, Liu SH, et al. Gene transfering mouse fetal brain in vivo［J］. Letters in Biotechnology, 2010, 21(3):413-415. (in Chinese)

吴燕,丁学锋, 刘淑红，等. 小鼠胎脑组织活体转基因技术的建立［J］. 生物技术通讯, 2010, 21(3):413-415.

［7］Zhang P, Wu CY, Tao QX, et al. Influence of three puncture sites of in utero electroporation on embryonic cortex development［J］. Chinese Journal of Nearotraumatic Surgery (Electronic Edition), 2015, 1 (4):27-34. (in Chinese)

张鹏，吴翠莹，陶庆霞，等. 不同穿刺点行子宫内电转对小鼠胚胎皮层发育的影响［J］. 中华神经创伤外科电子杂志, 2015, 1(4):27-34.

［8］Baumgart J, Baumgart N. Cortex-, hippocampus-, thalamus-, hypothalamus-, lateral septal nucleus- and striatum-specific in utero electroporation in the C57BL/6 mouse［J］. J Vis Exp, 2016, (107): e53303.

［9］Seuntjens E, Nityanandam A, Miquelajauregui A, et al. Sip1 regulates sequential fate decisions by feedback signaling from postmitotic neurons to progenitors［J］. Nat Neurosci, 2009, 12(11):1373-1380.

［10］Dominguez MH, Ayoub AE, Rakic P. POU-Ⅲ transcription factors (Brn1, Brn2, and Oct6) influence neurogenesis, molecular identity, and migratory destination of upper-layer cells of the cerebral cortex［J］. Cereb Cortex, 2013, 23(11):2632-2643.

［11］Walantus W, Castaneda D, Elias L, et al. In utero intraventricular injection and electroporation of E15 mouse embryos［J］. J Vis Exp,2007(6):e239.

［12］Haddad-Tovolli R, Szabo NE, Zhou X, et al. Genetic manipulation of the mouse developing hypothalamus through in utero electroporation［J］. J Vis Exp, 2013, (77): e50412.