MRC London Institute of Medical Sciences (LMS), London W12 0NN, UK.
Institute of Clinical Sciences (ICS), Imperial College, London W12 0NN, UK.
STAR Protoc. 2021 Aug 9;2(3):100726. doi: 10.1016/j.xpro.2021.100726. eCollection 2021 Sep 17.
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 Percharde et al. (2018).
LINE1 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。我们提出了一种在小鼠胚胎干细胞和胚胎中特异性敲低 LINE1 的方案，包括核转染和 LINE 反义寡核苷酸的受精卵显微注射的详细信息，随后进行 RNA FISH 验证。该方案可用于发育以及其他被认为表达 LINE1 的细胞类型。有关该方案使用和执行的完整信息，请参阅 Percharde 等人（2018 年）。
