Merck
CN

Depletion of nuclear LINE1 RNA in mouse ESCs and embryos.

STAR protocols (2021-08-20)
Michelle Percharde, Chih-Jen Lin, Miguel Ramalho-Santos
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
Sigma-Aldrich
Hyaluronidase from bovine testes, Type IV-S, powder, suitable for mouse embryo cell culture, 750-3000 units/mg solid
Sigma-Aldrich
Dextran sulfate sodium salt from Leuconostoc spp., for molecular biology, average Mw >500,000 (dextran starting material), contains 0.5-2% phosphate buffer