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Dev Biol 2021 Feb 01;470:74-83. doi: 10.1016/j.ydbio.2020.10.014.
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Trim-Away mediated knock down uncovers a new function for Lbh during gastrulation of Xenopus laevis.

Weir E , McLinden G , Alfandari D , Cousin H .

We previously identified the protein Lbh as necessary for cranial neural crest (CNC) cell migration in Xenopus through the use of morpholinos. However, Lbh is a maternally deposited protein and morpholinos achieve knockdowns through prevention of translation. In order to investigate the role of Lbh in earlier embryonic events, we employed the new technique "Trim-Away" to degrade this maternally deposited protein. Trim-Away utilizes the E3 ubiquitin ligase trim21 to degrade proteins targeted with an antibody and was developed in mammalian systems. Our results show that Xenopus is amenable to the Trim-Away technique. We also show that early knockdown of Lbh in Xenopus results in defects in gastrulation that present with a decrease in fibronectin matrix assembly, an increased in mesodermal cell migration and decrease in endodermal cell cohesion. We further show that the technique is also effective on a second abundant maternal protein PACSIN2. We discuss potential advantages and limit of the technique in Xenopus embryos as well as the mechanism of gastrulation inhibition.

PubMed ID: 33159936
PMC ID: PMC7855437
Article link: Dev Biol
Grant support: [+]

Species referenced: Xenopus laevis
Genes referenced: fn1 lbh pacsin2
GO keywords: ubiquitin ligase complex [+]

Article Images: [+] show captions
References [+] :
Briegel, Congenital heart disease reminiscent of partial trisomy 2p syndrome in mice transgenic for the transcription factor Lbh. 2005, Pubmed