Researchers adapted a CRISPR-Cas9 system to CRKP to elucidate how mutations in clinical CRKP isolates induce polymyxin resistance (PR). They demonstrated that CrrB was a positive regulator of PR, and common clinical mutations lead to the addition of both 4-amino-4-deoxy-L-arabinose and phosophethanolamine to lipid A.
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McConville, T. H., Annavajhala, M. K., Giddins, M. J., Macesic, N., Herrera, C. M., Rozenberg, F. D., Bhushan, G. L., Ahn, D., Mancia, F., Trent, M. S., & Uhlemann, A.-C. (2020). CrrB Positively Regulates High-Level Polymyxin Resistance and Virulence in Klebsiella pneumoniae. Cell Reports, 33(4). https://doi.org/10.1016/j.celrep.2020.108313 Cite
Researchers performed a features-oriented CRISPR-utilized systematic screen of OCT4-bound cis-regulatory elements using CRISPR-Cas9 to identify functional enhancers important for pluripotency maintenance in mouse embryonic stem cells.
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Wang, H. F., Warrier, T., Farran, C. A., Zheng, Z. H., Xing, Q. R., Fullwood, M. J., Zhang, L.-F., Li, H., Xu, J., Lim, T.-M., & Loh, Y.-H. (2020). Defining Essential Enhancers for Pluripotent Stem Cells Using a Features-Oriented CRISPR-Cas9 Screen. Cell Reports, 33(4). https://doi.org/10.1016/j.celrep.2020.108309 Cite
Scientists describe the T-cell Optimized for Packaging (TOP) vector for delivering guide RNAs and transgenes into primary T cells
[Molecular Therapy-Methods & Clinical Development]
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Investigators used CRISPR/Cas9 gene editing technology in human (h)PSCs to create a unique suite of four isogenic homozygous variants at amino acid positions 16(G/R) and 27(G/Q), which reside in the N-terminus of the β2AR.
[Molecular Therapy-Methods & Clinical Development]
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Kondrashov, A., Yusof, N. A. N. M., Hasan, A., Goulding, J., Kodagoda, T., Hoang, D. M., Vo, N. T. N., Melarangi, T., Dolatshad, N., Gorelik, J., Hill, S., Harding, S. E., & Denning, C. (2020). CRISPR/Cas9 mediated generation and analysis of N-terminus polymorphic models of beta2-adrenoreceptor in isogenic hPSC derived cardiomyocytes. Molecular Therapy - Methods & Clinical Development, 0(0). https://doi.org/10.1016/j.omtm.2020.10.019 Cite
EdiGene, Inc. announced the Center for Drug Evaluation of China National Medical Products Administration has accepted for review the company’s Investigational New Drug application for ET-01, autologous CD34+ hematopoietic stem/progenitor cells with the erythroid -specific enhancer of the BCL11A gene modified by CRISPR/Cas9, an investigational gene-editing therapy for patients with transfusion dependent β-thalassemia.
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Scientists present a microfluidic continuous-flow electrotransfection device designed for precise, consistent, and high-throughput genetic modification of target cells in cellular therapy manufacturing applications.
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High-throughput continuous-flow microfluidic electroporation of mRNA into primary human T cells for applications in cellular therapy manufacturing | Scientific Reports. (n.d.). Retrieved October 22, 2020, from https://www.nature.com/articles/s41598-020-73755-0 Cite
By using a high-throughput CRISPR/Cas9-based genetic screen in HepG2 hepatocellular carcinoma cells to search for metabolic proteins inhibiting ferroptosis, researchers identified a branched-chain amino acid aminotransferase 2 as a novel suppressor of ferroptosis.
[Cell Death & Differentiation]
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Wang, K., Zhang, Z., Tsai, H., Liu, Y., Gao, J., Wang, M., Song, L., Cao, X., Xu, Z., Chen, H., Gong, A., Wang, D., Cheng, F., & Zhu, H. (2020). Branched-chain amino acid aminotransferase 2 regulates ferroptotic cell death in cancer cells. Cell Death & Differentiation, 1–15. https://doi.org/10.1038/s41418-020-00644-4 Cite
Host rejection driven by HLA disparity in adoptively transferred allogeneic T cells remains a key obstacle to the universal donor T cell therapy. To evade donor HLA-mediated immune rejection, scientists attempted to eliminate T cell’s HLA through the CRISPR/Cas9 gene editing system.
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Using CRISPR/Cas9-mediated genome editing, scientists established HEK293 cells with knockout of GRK2, GRK3 or both to dissect their individual contributions in β-arrestin2 recruitment and μ-OR internalization upon stimulation with four different agonists.
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Dissecting the roles of GRK2 and GRK3 in μ-opioid receptor internalization and β-arrestin2 recruitment using CRISPR/Cas9-edited HEK293 cells | Scientific Reports. (n.d.). Retrieved October 16, 2020, from https://www.nature.com/articles/s41598-020-73674-0 Cite
Gene correction using CRISPR-Cas9 is an extension of gene therapy that has received considerable attention in recent years and boasts many possible uses beyond classical gene therapy approaches.
Investigators exploited the DMP1 promoter as a predictive marker of MSC differentiation towards osteoblasts. Using CRISPR/Cas9 technology, they identified a distinctive change in fluorescence intensities of GFP knock‐in and osteoblast differentiation.
[Journal of Gene Medicine]
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Shahabipour, F., Oskuee, R. K., Shokrgozar, M. A., Naderi‐Meshkin, H., Goshayeshi, L., & Bonakdar, S. (n.d.). CRISPR/Cas9 mediated GFP-Human Dentin Matrix Protein 1 (DMP1) Promoter knock-in at the ROSA26 locus in mesenchymal stem cell for monitoring osteoblast differentiation. The Journal of Gene Medicine, n/a(n/a), e3288. https://doi.org/10.1002/jgm.3288 Cite