Morpholinos Do Not Elicit an Innate Immune Response during Early Xenopus Embryogenesis

Dev Cell. 2019 May 20;49(4):643-650.e3. doi: 10.1016/j.devcel.2019.04.019.

Abstract

It has recently been reported that a common side effect of translation-blocking morpholino antisense oligonucleotides is the induction of a set of innate immune response genes in Xenopus embryos and that splicing-blocking morpholinos lead to unexpected off-target mis-splicing events. Here, we present an analysis of all publicly available Xenopus RNA sequencing (RNA-seq) data in a reexamination of the effects of translation-blocking morpholinos on the innate immune response. Our analysis does not support the authors' general conclusion, which was based on a limited number of RNA-seq datasets. Moreover, the strong induction of an immune response appears to be specific to the tbxt/tbxt2 morpholinos. The more comprehensive study presented here indicates that using morpholinos for targeted gene knockdowns remains of considerable value for the rapid identification of gene function.

Keywords: Xenopus; brachyury; gene knockdown; innate immune response; loss of function; reverse genetics; tbxt; translation blocking morpholino; zebrafish.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Review

MeSH terms

  • Animals
  • Embryonic Development / drug effects
  • Gene Knockdown Techniques
  • Immunity, Innate / immunology*
  • Immunity, Innate / physiology
  • Morpholinos / immunology*
  • Morpholinos / metabolism*
  • Oligonucleotides, Antisense / genetics
  • RNA Splicing
  • T-Box Domain Proteins / genetics
  • T-Box Domain Proteins / metabolism
  • Transcriptome / genetics
  • Xenopus / embryology
  • Xenopus / genetics
  • Xenopus Proteins / genetics
  • Xenopus Proteins / metabolism
  • Xenopus laevis / embryology
  • Xenopus laevis / genetics

Substances

  • Morpholinos
  • Oligonucleotides, Antisense
  • T-Box Domain Proteins
  • TBXT protein, Xenopus
  • Xenopus Proteins