CRISPR editing of the GLI1 first intron abrogates GLI1 expression and differentially alters lineage commitment
Rattachement africain : us, ru. Niveau de preuve : code pays fourni par la source.
Le résumé fourni par la source
GLI1 is one of three GLI family transcription factors that mediate Sonic Hedgehog signaling, which plays a role in development and cell differentiation. GLI1 forms a positive feedback loop with GLI2 and likely with itself. To determine the impact of GLI1 and its intronic regulatory locus on this transcriptional loop and human stem cell differentiation, we deleted the region containing six GLI binding sites in the human GLI1 intron using CRISPR/Cas9 editing to produce H1 human embryonic stem cell (hESC) GLI1-edited clones. Editing out this intronic region, without removing the entire GLI1 gene, allowed us to study the effects of this highly complex region, which binds transcription factors in a variety of cells. The roles of GLI1 in human ESC differentiation were investigated by comparing RNA sequencing, quantitative-real time PCR (q-rtPCR), and functional assays. Editing this region resulted in GLI1 transcriptional knockdown, delayed neural commitment, and inhibition of endodermal and mesodermal differentiation during spontaneous and directed differentiation experiments. We found a delay in the onset of early osteogenic markers, a reduction in the hematopoietic potential to form granulocyte units, and a decrease in cancer-related gene expression. Furthermore, inhibition of GLI1 via antagonist GANT-61 had similar in vitro effects. These results indicate that the GLI1 intronic region is critical for the feedback loop and that GLI1 has lineage-specific effects on hESC differentiation. Our work is the first study to document the extent of GLI1 abrogation on early stages of human development and to show that GLI1 transcription can be altered in a therapeutically useful way.
Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.
Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- CRISPR editing of the GLI1 first intron abrogates GLI1 expression and differentially alters lineage commitment
- Date Crossref
- 22/02/2021
- Éditeur
- Oxford University Press (OUP)
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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Northwestern University Ann & Robert H. Lurie Children's Hospital of Chicago pays non établi dans la noticeUniversité ou école supérieure
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Lurie Children's Hospital pays non établi dans la noticeÉtablissement de santé
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Russian Academy of Sciences pays non établi dans la noticeOrganisme public
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Institute of Theoretical and Experimental Biophysics pays non établi dans la noticeStructure de recherche
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Stanley Manne Children's Research Institute Developmental Biology Program pays non établi dans la noticeStructure de recherche
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ARTEC Biotech Inc pays non établi dans la noticeEntreprise
Ann & Robert H. Lurie Children's Hospital of Chicago — Northwestern University, Lurie Children's Hospital et Russian Academy of Sciences, avec 3 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.