Journal: Scientific reports

Article Title: CRISPR/Cas9-mediated deletion of a GA-repeat in human GPM6B leads to disruption of neural cell differentiation from NT2 cells.

doi: 10.1038/s41598-024-52675-3

Figure Lengend Snippet: Figure 1. In-silico analysis of GPM6B expression. (a) Across several primates, the expression of GPM6B was at its highest level in the human brain. (b) Our analysis of the RNA-seq datasets retrieved from GEO showed a positive correlation between GPM6B and neural cell differentiation markers, such as GFAP, TUBB3, and MAP2, after differentiation of NT2 cells into neural cells (21 days under RA treatment). HUM, human; CHP, chimpanzee; OWM, Old-World monkeys; NWM, New-World monkeys; MLM, mouse lemur.

Article Snippet: NTERA-2 (NT2) cell line (Cat. #ACC-527, RRID:CVCL_0034; was purchased from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH), resembling characteristics of human neuronal progenitor cell, Figure 4.

Techniques: In Silico, Expressing, RNA Sequencing, Cell Differentiation

Journal: Scientific reports

Article Title: CRISPR/Cas9-mediated deletion of a GA-repeat in human GPM6B leads to disruption of neural cell differentiation from NT2 cells.

doi: 10.1038/s41598-024-52675-3

Figure Lengend Snippet: Figure 3. Measuring the expression level of the edited GPM6B gene at the RNA and protein levels in NT2 cells. (a) The expression level of GPM6B was evaluated in the untreated and edited NT2 pool cells, using qRT-PCR. The expression level of GPM6B decreased significantly in the edited pool cells (p < 0.05). (b) The expression of GPM6B was assessed in the isolated single clones, using qRT-PCR. The expression of GPM6B was significantly decreased in the C1 cells, compared to the untreated and the SC3 cells. (c) Western blotting assay confirmed that the expression level of GPM6B was decreased in the C1 cells more efficiently, compared to the untreated and SC3 cells. The original blot images are presented in Supplementary Fig. Xd. (d) the predicted TF binding sites at GA-repeat site and its flanking sequence, is presented based on JASPAR CORE 2022 and ChIP-seq data from ENCODE. The GA-repeat site is highlighted with light blue.

Article Snippet: NTERA-2 (NT2) cell line (Cat. #ACC-527, RRID:CVCL_0034; was purchased from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH), resembling characteristics of human neuronal progenitor cell, Figure 4.

Techniques: Expressing, Quantitative RT-PCR, Isolation, Clone Assay, Western Blot, Binding Assay, Sequencing, ChIP-sequencing

Journal: Scientific reports

Article Title: CRISPR/Cas9-mediated deletion of a GA-repeat in human GPM6B leads to disruption of neural cell differentiation from NT2 cells.

doi: 10.1038/s41598-024-52675-3

Figure Lengend Snippet: Figure 5. Disrupted differentiation of NT2 cells to astrocytic and neural cells as a result of GA-repeat deletion and GPM6B downregulation. Deletion of the GA-repeat in the regulatory region of GPM6B decreased the expression of this gene. Consequently, the number of differentiated cells expressing GFAP, TUBB3, and MAP2 decreased significantly in the C1 compared to the SC3 cells.

Article Snippet: NTERA-2 (NT2) cell line (Cat. #ACC-527, RRID:CVCL_0034; was purchased from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH), resembling characteristics of human neuronal progenitor cell, Figure 4.

Techniques: Expressing