In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9

Swiech, Lukasz; Heidenreich, Matthias; Banerjee, Abhishek; Habib, Naomi; Li, Yinqing; Trombetta, John J.; Sur, Mriganka; Zhang, Feng

doi:10.1038/nbt.3055

Cited by 693 publications

(671 citation statements)

References 41 publications

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“…Recently, SpCas9 has been proven to be a powerful tool for making precise genomic perturbations in vivo. It has been used in vivo to study the function of genes in the liver and brain [42,43] and to cure metabolic and genetic diseases in animal models [44]. Here, we demonstrated that SpCas9 can be used to disrupt the genes in adult mouse islets.…”

Section: Discussionmentioning

confidence: 93%

RNA-binding protein CUGBP1 regulates insulin secretion via activation of phosphodiesterase 3B in mice

Zhai

Yang

et al. 2016

Diabetologia

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Aims/hypothesis CUG-binding protein 1 (CUGBP1) is a multifunctional RNA-binding protein that regulates RNA processing at several stages including translation, deadenylation and alternative splicing, as well as RNA stability. Recent studies indicate that CUGBP1 may play a role in metabolic disorders. Our objective was to examine its role in endocrine pancreas function through gain-and loss-of-function experiments and to further decipher the underlying molecular mechanisms. Methods A mouse model in which type 2 diabetes was induced by a high-fat diet (HFD; 60% energy from fat) and mice on a standard chow diet (10% energy from fat) were compared. Pancreas-specific CUGBP1 overexpression and knockdown mice were generated. Different lengths of the phosphodiesterase subtype 3B (PDE3B) 3′ untranslated region (UTR) were cloned for luciferase reporter analysis. Purified CUGBP1 protein was used for gel shift experiments.Results CUGBP1 is present in rodent islets and in beta cell lines; it is overexpressed in the islets of diabetic mice. Compared with control mice, the plasma insulin level after a glucose load was significantly lower and glucose clearance was greatly delayed in mice with pancreas-specific CUGBP1 overexpression; the opposite results were obtained upon pancreas-specific CUGBP1 knockdown. Glucose-and glucagon-like peptide1 (GLP-1)-stimulated insulin secretion was significantly attenuated in mouse islets upon CUGBP1 overexpression. This was associated with a strong decrease in intracellular cAMP levels, pointing to a potential role for cAMP PDEs. CUGBP1 overexpression had no effect on the mRNA levels of PDE1A, 1C, 2A, 3A, 4A, 4B, 4D, 7A and 8B subtypes, but resulted in increased PDE3B expression. CUGBP1 was found to directly bind to a specific ATTTGTT sequence residing in the 3′ UTR of PDE3B and stabilised PDE3B mRNA. In the presence of the PDE3 inhibitor cilostamide, glucose-and GLP-1-stimulated insulin secretion was no longer reduced by CUGBP1 overexpression. Similar to CUGBP1, PDE3B was overexpressed in the islets of diabetic mice. Conclusions/interpretation We conclude that CUGBP1 is a critical regulator of insulin secretion via activating PDE3B. Repressing this protein might provide a potential strategy for treating type 2 diabetes.

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Section: Discussionmentioning

confidence: 93%

RNA-binding protein CUGBP1 regulates insulin secretion via activation of phosphodiesterase 3B in mice

Zhai

Yang

et al. 2016

Diabetologia

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“…There are two major roadblocks for building these kinds of cellular models. The first one concerns gene editing and, over the past 3 years, has become largely historical: Until recently, genome engineering in human stem cells and neurons has been challenging but transfection of CRISPR plasmids or ribonucleoproteins provides an easy, efficient technique for engineering human cells (Peters et al 2008;Swiech et al 2015). The second major hurdle has been neural differentiation.…”

Section: Bottom-up Approaches Using Exome Sequencing In Autismmentioning

confidence: 99%

Multiscale Genome Engineering: Genome-Wide Screens and Targeted Approaches

Sanjana

2017

Research and Perspectives in Neurosciences

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“…This powerful technology facilitates targeted DNA double-strand breaks at specific sites in the mammalian genome and takes advantage of the non-homologous end joining (NHEJ) to introduce insertions or deletions (indels). [13][14][15] We demonstrate that miR-10b gene editing is deleterious for all glioma cells and GBM-initiating stem-like cells (GSCs) studied, as their viability strictly depends on miR-10b expression. Furthermore, we show that lentivirus-mediated miR-10b editing with CRISPR-Cas9 strongly impairs the growth of orthotopic GBM in mice, supporting targeted miR-10b gene editing as a promising therapeutic approach for GBM.…”

Section: Introductionmentioning

confidence: 99%