Using CRISPR/Cas9 genome editing, scientists obtaineded SH-SY5Y cell lines with frameshift mutations on one or both SHANK2 alleles. They investigated the effects of the different SHANK2 mutations on cell morphology, cell proliferation and differentiation potential during early neuronal differentiation.
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Unsicker, C., Cristian, F.-B., von Hahn, M., Eckstein, V., Rappold, G. A., & Berkel, S. (2021). SHANK2 mutations impair apoptosis, proliferation and neurite outgrowth during early neuronal differentiation in SH-SY5Y cells. Scientific Reports, 11(1), 2128. https://doi.org/10.1038/s41598-021-81241-4 Cite
The authors identified a novel SphK1-targeting microRNA, microRNA-6784 (miR-6784). They showed that miR-6784 was located at the cytoplasm of A431 skin squamous cell carcinoma cells.
The authors identified hypotaurine as one of the top-ranked metabolites for differentiating low- and high-grade tumors, and that there was also a strong association between the levels of intratumoral hypotaurine and expression of its biosynthetic enzyme, cysteamine (2-aminoethanethiol) dioxygenase.
[Cell Death Discovery]
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Researchers performed electroporation of CD34+ hematopoietic stem and progenitor cells obtained from healthy donors, with CRISPR-Cas9 targeting the BCL11A erythroid-specific enhancer.
[New England Journal of Medicine]
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Frangoul, H., Altshuler, D., Cappellini, M. D., Chen, Y.-S., Domm, J., Eustace, B. K., Foell, J., Fuente, J. de la, Grupp, S., Handgretinger, R., Ho, T. W., Kattamis, A., Kernytsky, A., Lekstrom-Himes, J., Li, A. M., Locatelli, F., Mapara, M. Y., Montalembert, M. de, Rondelli, D., … Corbacioglu, S. (2020). CRISPR-Cas9 Gene Editing for Sickle Cell Disease and β-Thalassemia. New England Journal of Medicine. https://doi.org/10.1056/NEJMoa2031054 Cite
Autophagosome formation and autophagic flux were assessed by evaluating endogenous LC3-II levels and ectopic expression of EGFP-LC3 and mRFP-EGFP-LC3 in breast cancer cells.
[British Journal of Cancer]
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The authors describe Tracking Recombination Alleles in Clonal Engraftment using sequencing (TRACE-Seq), a methodology that utilizes barcoded AAV6 donor template libraries, carrying in-frame silent mutations or semi-randomized nucleotides outside the coding region, to track the in vivo lineage contribution of gene-targeted hematopoietic stem and progenitor cell clones.
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Sharma, R., Dever, D. P., Lee, C. M., Azizi, A., Pan, Y., Camarena, J., Köhnke, T., Bao, G., Porteus, M. H., & Majeti, R. (2021). The TRACE-Seq method tracks recombination alleles and identifies clonal reconstitution dynamics of gene targeted human hematopoietic stem cells. Nature Communications, 12(1), 472. https://doi.org/10.1038/s41467-020-20792-y Cite
CRISPR/Cas9-mediated ARID1A knockout in primary TP53-/- human gastric organoids induced morphologic dysplasia, tumorigenicity and mucinous differentiation.
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Lo, Y.-H., Kolahi, K. S., Du, Y., Chang, C.-Y., Krokhotin, A., Nair, A., Sobba, W. D., Karlsson, K., Jones, S. J., Longacre, T. A., Mah, A. T., Tercan, B., Sockell, A., Xu, H., Seoane, J. A., Chen, J., Shmulevich, I., Weissman, J. S., Curtis, C., … Kuo, C. J. (2021). A CRISPR/Cas9-engineered ARID1A-deficient human gastric cancer organoid model reveals essential and non-essential modes of oncogenic transformation. Cancer Discovery. https://doi.org/10.1158/2159-8290.CD-20-1109 Cite
The authors report a genome-wide loss-of-function screening that identified gene mutations that could significantly reduce the rate of self-diploidization in haploid ESCs.
[Stem Cell Reports]
The authors report an efficient, network-based drug-screening platform developed by integrating mathematical modeling and the pathological features of Alzheimer’s disease with human induced pluripotent stem cell-derived cerebral organoids, including CRISPR-Cas9-edited isogenic lines.
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Park, J.-C., Jang, S.-Y., Lee, D., Lee, J., Kang, U., Chang, H., Kim, H. J., Han, S.-H., Seo, J., Choi, M., Lee, D. Y., Byun, M. S., Yi, D., Cho, K.-H., & Mook-Jung, I. (2021). A logical network-based drug-screening platform for Alzheimer’s disease representing pathological features of human brain organoids. Nature Communications, 12(1), 280. https://doi.org/10.1038/s41467-020-20440-5 Cite
The authors conducted two lines of genome-editing experiments of mouse hematopoietic stem cells (HSCs) with the clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9).
[Molecular Therapy-Methods & Clinical Development]
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Researchers used CRISPR/Cas9 gene editing to engineer an NRL-deficient embryonic stem cell line, and differentiated it into retinal organoids.
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NRL−/− gene edited human embryonic stem cells generate rod‐deficient retinal organoids enriched in S‐cone‐like photoreceptors - Cuevas - - STEM CELLS - Wiley Online Library. (n.d.). Retrieved January 8, 2021, from https://stemcellsjournals.onlinelibrary.wiley.com/doi/abs/10.1002/stem.3325?af=R Cite