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Mech Dev August 1, 2017; 146 1-9.
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An analysis of MyoD-dependent transcription using CRISPR/Cas9 gene targeting in Xenopus tropicalis embryos.

McQueen C , Pownall ME .

Myogenic regulatory factors (MRFs) are known to have essential roles in both the establishment and differentiation of the skeletal muscle cell lineage. MyoD is expressed early in the Xenopus mesoderm where it is present and active several hours before the activation of muscle differentiation genes. Previous studies in cultured cells and in Xenopus laevis have identified sets of genes that require MyoD prior to differentiation of skeletal muscle. Here we report results from experiments using CRISPR/Cas9 to target the MyoD gene in the diploid frog Xenopus tropicalis, that are analysed by RNA-seq at gastrula stages. We further investigate our data using cluster analysis to compare developmental expression profiles with that of MyoD and α-cardiac actin, reference genes for skeletal muscle determination and differentiation. Our findings provide an assessment of using founder (F0) Xenopus embryos from CRISPR/Cas9 protocols for transcriptomic analyses and we conclude that although targeted F0 embryos are genetically mosaic for MyoD, there is significant disruption in the expression of a specific set of genes. We discuss candidate target genes in context of their role in the sub-programs of MyoD regulated transcription.

PubMed ID: 28536000
Article link: Mech Dev
Grant support: [+]

Species referenced: Xenopus tropicalis Xenopus laevis
Genes referenced: actc1 babam1 bmpr1b cdx1 decr2 dnajc24 epn1 flvcr2 foxc1 foxc2 fstl1 gbx2.2 gli2 herpud2 mrps30 msi1 myod1 nkx6-2 pam pbx2 pcdh8.2 pdlim7 pex16 pgk1 pgp pmm2 pygm rbm20 rbm24 rnf157 rnf7 slc13a4l sp5 sp8 tsfm zeb2

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