Depletion of nuclear LINE1 RNA in mouse ESCs and embryos.
Depletion of nuclear LINE1 RNA in mouse ESCs and embryos.
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DOI：
10.1016/j.xpro.2021.100726
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发表时间：
2021-09-17
期刊：
影响因子：
--
通讯作者：
Ramalho-Santos M
中科院分区：
文献类型：
--
作者：
Percharde M;Lin CJ;Ramalho-Santos M
LINE 1是最活跃和丰富的逆转录转座子家族;它与许多病理学以及早期胚胎发育有关。我们提出了一个协议，专门敲低LINE 1在小鼠胚胎干细胞和胚胎，包括核转染和受精卵显微注射的LINE反义寡核苷酸的细节，随后RNA FISH验证。该方案可用于开发，以及据信表达LINE 1的其他细胞类型。有关本方案使用和执行的完整信息，请参见。反义寡核苷酸（ASOs）核转染入ESC以特异性消除LINE 1 RNA受精卵显微注射LINE 1 ASOs以进行胚胎敲低LINE 1 RNA的可视化和通过RNA FISH验证消除LINE 1是最活跃和最丰富的逆转录转座子家族;它涉及许多病理学以及早期胚胎发育。我们提出了一种在小鼠胚胎干细胞和胚胎中特异性敲除LINE 1的方案，包括LINE反义寡核苷酸的核转染和受精卵显微注射的细节，然后进行RNA FISH验证。该方案可用于开发，以及据信表达LINE 1的其他细胞类型。
LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote microinjection of LINE antisense oligos, followed by RNA FISH validation. This protocol can be used in development, as well as other cell types where LINE1 is believed to be expressed. For complete information on the use and execution of this protocol, please refer to. Nucleofection of antisense oligos (ASOs) into ESCs for specific depletion of LINE1 RNA Zygote microinjection of LINE1 ASOs for embryonic knockdown Visualization of LINE1 RNA and validation of depletion by RNA FISH LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote microinjection of LINE antisense oligos, followed by RNA FISH validation. This protocol can be used in development, as well as other cell types where LINE1 is believed to be expressed.
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影响因子：
64.5
作者：
Percharde M;Lin CJ;Yin Y;Guan J;Peixoto GA;Bulut-Karslioglu A;Biechele S;Huang B;Shen X;Ramalho-Santos M
通讯作者：
Ramalho-Santos M
影响因子：
48
作者：
Raj, Arjun;van den Bogaard, Patrick;Rifkin, Scott A.;van Oudenaarden, Alexander;Tyagi, Sanjay
通讯作者：
Tyagi, Sanjay
影响因子：
64.8
作者：
Ying, Qi-Long;Wray, Jason;Smith, Austin
通讯作者：
Smith, Austin
影响因子：
11.8
作者：
Lin, Chih-Jen;Koh, Fong Ming;Wong, Priscilla;Conti, Marco;Ramalho-Santos, Miguel
通讯作者：
Ramalho-Santos, Miguel
影响因子：
4.6
作者：
Lin, Chih-Jen;Conti, Marco;Ramalho-Santos, Miguel
通讯作者：
Ramalho-Santos, Miguel