Blossom of CRISPR technologies and applications in disease treatment

Liu, Huayi; Wang, Lian; Luo, Yunzi

doi:10.1016/j.synbio.2018.10.003

Cited by 17 publications

(13 citation statements)

References 188 publications

(224 reference statements)

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“…Gene therapy has recently gained successes in treating various kinds of deafness [33, 113–118]. With the advances in gene therapy delivery and safety such as base editing [27, 119, 120] and improved encapsulation via nanoparticles or hydrogels [121–123], we are optimistic that treatment options for patients will become more available in the not too distant future. Meanwhile, chemicals that target the potassium channel activity also remain a promising therapeutic approach to reduce or delay the disease progression.…”

Section: Discussionmentioning

confidence: 99%

Hearing consequences in Gjb2 knock-in mice: implications for human p.V37I mutation

Lin

Zhang

et al. 2019

Aging

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Human p.V37I mutation of GJB2 gene was strongly correlated with late-onset progressive hearing loss, especially among East Asia populations. We generated a knock-in mouse model based on human p.V37I variant (c.109G>A) that recapitulated the human phenotype. Cochlear pathology revealed no significant hair cell loss, stria vascularis atrophy or spiral ganglion neuron loss, but a significant change in the length of gap junction plaques, which may have contributed to the observed mild endocochlear potential (EP) drop in homozygous mice lasting lifetime. The cochlear amplification in homozygous mice was compromised, but outer hair cells’ function remained unchanged, indicating that the reduced amplification was EP- rather than prestin-generated. In addition to ABR threshold elevation, ABR wave I latencies were also prolonged in aged homozygous animals. We found in homozygous IHCs a significant increase in ICa but no change in Ca2+ efficiency in triggering exocytosis. Environmental insults such as noise exposure, middle ear injection of KCl solution and systemic application of furosemide all exacerbated the pathological phenotype in homozygous mice. We conclude that this Gjb2 mutation-induced hearing loss results from 1) reduced cochlear amplifier caused by lowered EP, 2) IHCs excitotoxicity associated with potassium accumulation around hair cells, and 3) progression induced by environmental insults.

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Section: Discussionmentioning

confidence: 99%

Hearing consequences in Gjb2 knock-in mice: implications for human p.V37I mutation

Lin

Zhang

et al. 2019

Aging

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“…Considering the importance of gene regulation in understanding and manipulating cellular metabolism, development of robust and efficient gene regulation techniques is drawing increasing attention. CRISPRi and RNAi that function at the transcription and translation stages, respectively, are well established and widely used in biological research and disease treatment ( Laganà et al, 2014 ; Liu et al, 2018 ). With the advent of the type IV-D CRISPR effector (Cas13d), new programmable RNA-guided and RNA-targeting techniques are available, which have largely advanced the field of gene regulation ( Wang F. et al, 2019 ).…”

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas13d-Mediated Microbial RNA Knockdown

Zhang

Kang³

et al. 2020

Front. Bioeng. Biotechnol.

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RNA-guided and RNA-targeting type IV-D CRISPR/Cas systems (CRISPR/Cas13d) have recently been identified and employed for efficient and specific RNA knockdown in mammalian and plant cells. Cas13d possesses dual RNase activities and is capable of processing CRISPR arrays and cleaving target RNAs in a protospacer flanking sequence (PFS)-independent manner. These properties make this system a promising tool for multiplex gene expression regulation in microbes. Herein, we aimed to establish a CRISPR/Cas13d-mediated RNA knockdown platform for bacterial chassis. CasRx, Cas13d from Ruminococcus flavefaciens XPD3002, was selected due to its high activity. However, CasRx was found to be highly toxic to both Escherichia coli and Corynebacterium glutamicum, especially when it cooperated with its guide and target RNAs. After employing a low copy number vector, a tightly controlled promoter, and a weakened ribosome binding site, we successfully constructed an inducible expression system for CasRx and applied it for repressing the expression of a green fluorescent protein (GFP) in E. coli. Despite our efforts to optimize inducer usage, guide RNA (gRNA) architecture and combination, and target gene expression level, the highest gene repression efficiency was 30-50% at the protein level and ∼70% at the mRNA level. The moderate RNA knockdown is possibly caused by the collateral cleavage activity toward bystander RNAs, which acts as a mechanism of type IV-D immunity and perturbs microbial metabolism. Further studies on cellular response to CRISPR/Cas13d and improvement in RNA knockdown efficiency are required prior to practical application of this system in microbes.

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“…Luckily, with the advent of CRISPR systems, tools for site‐specific genome editing in Streptomyces were rapidly developed (Figure ). Those CRISPR‐based gene manipulation tools complement traditional approaches and facilitate discovery of new compounds with significant bioactivities . Ryan Cobb et al.…”

Section: Current Synthetic Biology Tools Developed For Applications Imentioning

confidence: 99%

“…Luckily, with the advent of CRISPR systems, tools for site-specific genome editing in Streptomyces were rapidly developed ( Figure 2). Those CRISPR-based gene manipulation tools complement traditional approaches and facilitate discovery of new compounds with significant bioactivities [19,23,24]. Ryan Cobb et al [25] introduced the CRISPR/Cas9 system into Streptomyces and achieved fast multiplex genome editing via selected gene knock-in and knock-out.…”

Section: In Vivo Genome Editing Tools In Streptomycesmentioning

confidence: 99%

Recent advances in natural products exploitation in Streptomyces via synthetic biology

Zhao

Wang

Luo

2019

Engineering in Life Sciences

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Natural products of microbial origin have proven to be the wellspring of clinically useful compounds for human therapeutics. Streptomyces species are predominant sources of bioactive compounds, most of which serve as potential drug candidates. While the exploitation of natural products has been severely reduced over the past two decades, the growing crisis of evolution and dissemination of drug resistant pathogens have again attracted great interest in this field. The emerging synthetic biology has been heralded as a new bioengineering platform to discover novel bioactive compounds and expand bioactive natural products diversity and production. Herein, we review recent advances in the natural products exploitation of Streptomyces with the applications of synthetic biology from three major aspects, including recently developed synthetic biology tools, natural products biosynthetic pathway engineering strategies as well as chassis host modifications.

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Blossom of CRISPR technologies and applications in disease treatment

Cited by 17 publications

References 188 publications

Hearing consequences in Gjb2 knock-in mice: implications for human p.V37I mutation

Hearing consequences in Gjb2 knock-in mice: implications for human p.V37I mutation

CRISPR/Cas13d-Mediated Microbial RNA Knockdown

Recent advances in natural products exploitation in Streptomyces via synthetic biology

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