Cornerstones of CRISPR–Cas in drug discovery and therapy

Fellmann, Christof; Gowen, Benjamin G.; Lin, Pei‐Chun; Doudna, Jennifer A.; Corn, Jacob E.

doi:10.1038/nrd.2016.238

Cited by 382 publications

(299 citation statements)

References 165 publications

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“…Directly targeting mutations into the NOD genetic background via CRISPR gene editing in NOD embryos is now possible 20 . Matching sgRNAs in exon 2 of Zbtb32 ( Figure 1A and B) and CAS9n were injected into 2 cell-stage embryos.…”

Section: Resultsmentioning

confidence: 99%

Loss of Zbtb32 in NOD mice does not significantly alter T cell responses.

et al. 2018

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Background : We previously identified the transcriptional regulator Zbtb32 as a factor that can promote T cell tolerance in the Non-Obese Diabetic (NOD) mouse, a model of Type 1 diabetes. Antigen targeted to DCIR2 + dendritic cells (DCs) in vivo inhibited both diabetes and effector T cell expansion in NOD mice. Furthermore, Zbtb32 was preferentially induced in autoreactive CD4 T cells stimulated by these tolerogenic DCIR2 + DCs, and overexpression of Zbtb32 in islet-specific T cells inhibited the diabetes development by limiting T cell proliferation and cytokine production. Methods : To further understand the role of Zbtb32 in T cell tolerance induction, we have now used CRISPR to target the Zbtb32 gene for deletion directly in NOD mice and characterized the mutant mice. We hypothesized that the systemic loss of Zbtb32 in NOD mice would lead to increased T cell activation and increased diabetes pathogenesis. Results : Although NOD.Zbtb32 -/- male NOD mice showed a trend towards increased diabetes incidence compared to littermate controls, the difference was not significant. Furthermore, no significant alteration in lymphocyte number or function was observed. Importantly, in vitro stimulation of lymphocytes from NOD.Zbtb32 -/- mice did not produce the expected hypersensitive phenotype observed in other genetic strains, potentially due to compensation by homologous genes. Conclusions : The loss of Zbtb32 in the NOD background does not result in the expected T cell activation phenotype.

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Section: Resultsmentioning

confidence: 99%

Loss of Zbtb32 in NOD mice does not significantly alter T cell responses.

et al. 2018

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“…Only recently discovered, CRISPR/Cas9 technology has already been enhanced to the point of fulfilling most of the genome editing and gene regulation currently demanded, ranging from the ability to perform multiple gene insertions, gene knockouts, combinatorial libraries, to advanced fine-tuning of biosynthetic pathways [23,39,47,95]. However, off-targeting remains an important limitation to the Retrotransposons are composed of similar DNA sequences, which ultimately allows to generate a promiscuous gRNA able to target several of these targets at once.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

Ferreira

David

Nielsen

2018

Journal of Industrial Microbiology and Biotechnology

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Clustered regularly interspaced short palindromic repeats (CRISPR) is poised to become one of the key scientific discoveries of the twenty-first century. Originating from prokaryotic and archaeal immune systems to counter phage invasions, CRISPRbased applications have been tailored for manipulating a broad range of living organisms. From the different elucidated types of CRISPR mechanisms, the type II system adapted from Streptococcus pyogenes has been the most exploited as a tool for genome engineering and gene regulation. In this review, we describe the different applications of CRISPR/Cas9 technology in the industrial biotechnology field. Next, we detail the current status of the patent landscape, highlighting its exploitation through different companies, and conclude with future perspectives of this technology.

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“…Moreover, the introduction of double-strand breaks by any method can be coupled with homology directed recombination (HDR) using DNA fragments or ssDNA oligonucleotides to introduce known allelic variations into specific sites in the genome. The consequence of these technological advances means that specific nucleotide changes can be engineered into any iPSC line to recapitulate the mutations causing any inborn error of hepatic metabolism no matter how rare 36 .…”

Section: Pluripotent Stem Cells As a Powerful Tool For Disease Modelingmentioning

confidence: 99%

“…The efficiency of using CRISPR/Cas9 coupled with homology directed repair is likely to substantially increase as new approaches are described, and existing techniques are optimized 36 . With advances in next generation sequencing, new allelic variations of uncertain significance are being identified that correlate with liver disease.…”

Section: Closing Remarksmentioning

confidence: 99%

Modeling Inborn Errors of Hepatic Metabolism Using Induced Pluripotent Stem Cells

Pournasr

Duncan

2017

ATVB

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Inborn errors of hepatic metabolism are due to deficiencies commonly within a single enzyme as a consequence of heritable mutations in the genome. Individually such diseases are rare, but collectively they are common. Advances in genome wide association studies (GWAS) and DNA sequencing have helped researchers identify the underlying genetic basis of such diseases. Unfortunately, cellular and animal models that accurately recapitulate these inborn errors of hepatic metabolism in the laboratory have been lacking. Recently investigators have exploited molecular techniques to generate induced pluripotent stem cells (iPSCs) from patients’ somatic cells. iPSCs can differentiate into a wide variety of cell types including hepatocytes thereby offering an innovative approach to unravel the mechanisms underlying inborn errors of hepatic metabolism. Moreover, such cell models could potentially provide a platform for the discovery of therapeutics. In this mini-review, we present a brief overview of the state-of-the-art in using pluripotent stem cells for such studies.

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Cornerstones of CRISPR–Cas in drug discovery and therapy

Cited by 382 publications

References 165 publications

Loss of Zbtb32 in NOD mice does not significantly alter T cell responses.

Loss of Zbtb32 in NOD mice does not significantly alter T cell responses.

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

Modeling Inborn Errors of Hepatic Metabolism Using Induced Pluripotent Stem Cells

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