bims-bicyki Biomed News
on Bicaudal-C1 and interactors in cystic kidney disease
Issue of 2021–01–10
fourteen papers selected by
Céline Gagnieux, École Polytechnique Fédérale de Lausanne
  1. Primary Cilia, Ciliogenesis and the Actin Cytoskeleton: A Little Less Resorption, A Little More Actin Please.
  2. Genetic and Sporadic Renal Cystic Diseases During the Perinatal and Neonatal Period: Core Curriculum 2021.
  3. Serum uromodulin levels, MR imaging findings, and their relationship with eGFR-based CKD staging in ADPKD patients.
  4. Dickkopf-3: Current Knowledge in Kidney Diseases.
  5. Suppression of Foxo3-Gatm by miR-132-3p Accelerates Cyst Formation by Up-Regulating ROS in Autosomal Dominant Polycystic Kidney Disease.
  6. [Targeting the ubiquitin system for treatment of cilia-related diseases].
  7. Activation of TRPV4 by mechanical, osmotic or pharmaceutical stimulation is anti-inflammatory blocking IL-1β mediated articular cartilage matrix destruction.
  8. Bi-allelic BRWD1 variants cause male infertility with asthenoteratozoospermia and likely primary ciliary dyskinesia.
  9. Host Antiviral Response Suppresses Ciliogenesis and Motile Ciliary Functions in the Nasal Epithelium.
  10. Functional inhibition of fatty acid binding protein 4 ameliorates impaired ciliogenesis in GCs.
  11. Ectoderm to mesoderm transition by down-regulation of actomyosin contractility.
  12. RhoA GTPase phosphorylated at tyrosine 42 by src kinase binds to β-catenin and contributes transcriptional regulation of vimentin upon Wnt3A.
  13. Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing.
  14. Gradual centriole maturation associates with the mitotic surveillance pathway in mouse development.
  1. Front Cell Dev Biol. 2020 ;8 622822
    Primary Cilia, Ciliogenesis and the Actin Cytoskeleton: A Little Less Resorption, A Little More Actin Please.
    Claire E L Smith, Alice V R Lake, Colin A Johnson.
      Primary cilia are microtubule-based organelles that extend from the apical surface of most mammalian cells, forming when the basal body (derived from the mother centriole) docks at the apical cell membrane. They act as universal cellular "antennae" in vertebrates that receive and integrate mechanical and chemical signals from the extracellular environment, serving diverse roles in chemo-, mechano- and photo-sensation that control developmental signaling, cell polarity and cell proliferation. Mutations in ciliary genes cause a major group of inherited developmental disorders called ciliopathies. There are very few preventative treatments or new therapeutic interventions that modify disease progression or the long-term outlook of patients with these conditions. Recent work has identified at least four distinct but interrelated cellular processes that regulate cilia formation and maintenance, comprising the cell cycle, cellular proteostasis, signaling pathways and structural influences of the actin cytoskeleton. The actin cytoskeleton is composed of microfilaments that are formed from filamentous (F) polymers of globular G-actin subunits. Actin filaments are organized into bundles and networks, and are attached to the cell membrane, by diverse cross-linking proteins. During cell migration, actin filament bundles form either radially at the leading edge or as axial stress fibers. Early studies demonstrated that loss-of-function mutations in ciliopathy genes increased stress fiber formation and impaired ciliogenesis whereas pharmacological inhibition of actin polymerization promoted ciliogenesis. These studies suggest that polymerization of the actin cytoskeleton, F-actin branching and the formation of stress fibers all inhibit primary cilium formation, whereas depolymerization or depletion of actin enhance ciliogenesis. Here, we review the mechanistic basis for these effects on ciliogenesis, which comprise several cellular processes acting in concert at different timescales. Actin polymerization is both a physical barrier to both cilia-targeted vesicle transport and to the membrane remodeling required for ciliogenesis. In contrast, actin may cause cilia loss by localizing disassembly factors at the ciliary base, and F-actin branching may itself activate the YAP/TAZ pathway to promote cilia disassembly. The fundamental role of actin polymerization in the control of ciliogenesis may present potential new targets for disease-modifying therapeutic approaches in treating ciliopathies.
    Keywords:  ROCK inhibitors; actin cytoskeleton; cilia; ciliogenesis; ciliopathies; cytoskeletal drugs; drug screen; polycystic kidney disease
    DOI:  https://doi.org/10.3389/fcell.2020.622822
  2. Am J Kidney Dis. 2021 Jan 05. pii: S0272-6386(20)31194-X. [Epub ahead of print]
    Genetic and Sporadic Renal Cystic Diseases During the Perinatal and Neonatal Period: Core Curriculum 2021.
    Rupesh Raina, Ronith Chakraborty, Sidharth K Sethi, Deepak Kumar, Kelly Gibson, Carsten Bergmann.
      Renal cystic disease encompasses a large variety of illnesses with variable phenotypic expression that can manifest in utero, in infancy, and in childhood. These diseases may be unilateral or bilateral and present with single or multiple cysts. Various cystic diseases may also progress to chronic kidney disease (CKD), hepatic disease, or kidney failure, potentially being life threatening. Due to the prevalence and life-altering complications of cystic kidney diseases in the pediatric population, it is crucial for clinicians and healthcare providers to pre-emptively detect and provide effective management. This installment of AJKD's Core Curriculum in Nephrology discusses the various genetic and sporadic kidney cystic diseases including multicystic dysplastic kidney, nephronophthisis, cystic dysplasia, hepatocyte nuclear factor 1-beta (HNF1-beta) nephropathy, Bardet-Biedl syndrome, Meckel-Gruber syndrome, Zellweger syndrome, calyceal diverticulum, autosomal recessive polycystic kidney disease (ARPKD), and autosomal dominant polycystic kidney disease (ADPKD). For reach of these renal cystic diseases, this article depicts the epidemiology, genetics and pathophysiology, diagnosis, presentation, and management, with emphasis on prenatal care and pregnancy counseling.
    Keywords:  Bardet-Biedl syndrome (BBS); HNF1-beta nephropathy; Zellweger spectrum disorders; calyceal diverticula (CD); children; ciliopathy; congenital disease; cystic dysplasia; fetal kidney anomaly; infantile nephronophthisis (NPHP); kidney disease; multicystic dysplastic kidney (MCDK); pediatric; polycystic kidney disease (PKD); prenatal; prenatal counseling; renal cystic disease; review
    DOI:  https://doi.org/10.1053/j.ajkd.2020.10.021
  3. Int Urol Nephrol. 2021 Jan 03.
    Serum uromodulin levels, MR imaging findings, and their relationship with eGFR-based CKD staging in ADPKD patients.
    Hale Nur Cansever, Funda Sari, Can Cevikol, Ramazan Cetinkaya, Gultekin Süleymanlar, Fevzi Ersoy.
       PURPOSE: Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disease that may progress to end-stage renal disease, characterized by increased kidney volume due to cystic formations. In this study, we aimed to investigate the relationship between serum uromodulin levels, total kidney volume and estimated glomerular filtration rate (eGFR) in patients with ADPKD.
    METHODS: This study included a total of 54 ADPKD patients and 18 healthy volunteers (control group). Total kidney volumes were calculated through magnetic resonance images using ellipsoid method. Serum uromodulin measurements were measured using an ELISA method.
    RESULTS: Serum uromodulin levels were lower in patients compared with the control group (2.47 ± 0.16 vs 2.6 ± 0.28, p = 0.021). There was no significant difference in uromodulin values among the patients in chronic kidney disease (CKD) stages 1-2, 3 and 4-5. TKV measurements of CKD stage 4-5 patients were significantly higher than the stage 1-2 patients (p = 0.015). A negative correlation was observed between TKV and eGFR (r = - 0.433, p = 0.001). A positive correlation was observed between uromodulin and eGFR (r = 0.274, p = 0.02). When the serum levels of uromodulin and the level of eGFR were evaluated using simple linear regression analysis, R2 value was found to be 0.075, suggesting that 7.5% change in serum uromodulin values corresponds with the change in eGFR value.
    CONCLUSION: These findings are consistent with previous studies that reported that serum uromodulin may be a good biomarker for demonstrating renal function in the early stages of CKD, before eGFR levels deteriorate. Serum uromodulin level may be useful in demonstrating renal functions in the follow-up of individuals with ADPKD.
    Keywords:  ADPKD; CKD; Total kidney volume; Uromodulin
    DOI:  https://doi.org/10.1007/s11255-020-02730-5
  4. Front Physiol. 2020 ;11 533344
    Dickkopf-3: Current Knowledge in Kidney Diseases.
    Xiangdong Fang, Jing Hu, Yanxia Chen, Wen Shen, Ben Ke.
      Dickkopf-related protein 3 (DKK3) is a secreted glycoprotein that has been implicated in the pathogenesis of a variety of diseases. Recent evidence suggests that urinary DKK3 may serve as a potential biomarker for monitoring kidney disease progression and assessing the effects of interventions. We review the biological role of DKK3 as an agonist in chronic kidney disease (CKD) and autosomal dominant polycystic kidney disease (ADPKD) and as an antagonist in idiopathic membranous nephropathy (IMN). In addition, we present the clinical applications of DKK3 in acute kidney disease and tubulointerstitial fibrosis, suggesting that urine DKK3 may be a potential biomarker for acute kidney disease and CKD. Further research into the mechanism of DKK3 and its use as a diagnostic tool, alone or in combination with other biomarkers, could prove clinically useful for better understanding the pathology of kidney diseases and improving early detection and treatment.
    Keywords:  Dickkopf-related protein 3; biomarker; chronic kidney disease; idiopathic membranous nephropathy; wnt signaling pathway
    DOI:  https://doi.org/10.3389/fphys.2020.533344
  5. Biomol Ther (Seoul). 2021 Jan 07.
    Suppression of Foxo3-Gatm by miR-132-3p Accelerates Cyst Formation by Up-Regulating ROS in Autosomal Dominant Polycystic Kidney Disease.
    Seonju Choi, Do Yeon Kim, Yejin Ahn, Eun Ji Lee, Jong Hoon Park.
      Accumulation of reactive oxygen species (ROS) is associated with the development of various diseases. However, the molecular mechanisms underlying oxidative stress that lead to such diseases like autosomal dominant polycystic kidney disease (ADPKD) remain unclear. Here, we observed that oxidative stress markers were increased in Pkd1f/f:HoxB7-Cre mice. Forkhead transcription factors of the O class (FOXOs) are known key regulators of the oxidative stress response, which have been observed with the expression of FoxO3a in an ADPKD mouse model in the present study. An integrated analysis of two datasets for differentially expressed miRNA, such as miRNA sequencing analysis of Pkd1 conditional knockout mice and microarray analysis of samples from ADPKD patients, showed that miR-132-3p was a key regulator of FOXO3a in ADPKD. miR-132-3p was significantly upregulated in ADPKD which directly targeted FOXO3 in both mouse and human cell lines. Interestingly, the mitochondrial gene Gatm was downregulated in ADPKD which led to a decreased inhibition of FOXO3. Overexpression of miR-132-3p coupled with knockdown of FOXO3 and Gatm increased ROS and accelerated cyst formation in 3D culture. This study reveals a novel mechanism involving miR-132-3p, FOXO3, and Gatm that is associated with the oxidative stress that occurs during cystogenesis in ADPKD.
    Keywords:  ADPKD; Cystogenesis; FOXO3; MicroRNA; ROS
    DOI:  https://doi.org/10.4062/biomolther.2020.197
  6. Nihon Yakurigaku Zasshi. 2021 ;156(1): 4-8
    [Targeting the ubiquitin system for treatment of cilia-related diseases].
    Yuhei Nishimura, Masaki Inagaki.
      The ubiquitin system regulates a wide variety of cellular functions. Not surprisingly, dysregulation of the ubiquitin system is associated with various disorders. Therefore, drugs that can modulate the functions of the ubiquitin system have been actively developed to treat these disorders. Chemical knockdown of pathogenic proteins using the ubiquitin-proteasome system is also a promising approach. The ubiquitin system regulates the assemble and disassemble of primary cilia through balanced control over the ubiquitination and deubiquitination of ciliary proteins. Primary cilia are antenna-like structures present in many vertebrate cells that sense and transduce extracellular cues to control cellular processes such as proliferation and differentiation. Impairment of primary cilia is associated with many diseases, including cancer and ciliopathy, a group of multisystem developmental disorders. In this review, we focus on the role of the ubiquitin system on cilia-related disorders and discuss the possibility of the ubiquitin system as therapeutic targets for these diseases through regulation of primary cilia formation.
    DOI:  https://doi.org/10.1254/fpj.20072
  7. Osteoarthritis Cartilage. 2021 Jan;pii: S1063-4584(20)31109-2. [Epub ahead of print]29(1): 89-99
    Activation of TRPV4 by mechanical, osmotic or pharmaceutical stimulation is anti-inflammatory blocking IL-1β mediated articular cartilage matrix destruction.
    S Fu, H Meng, S Inamdar, B Das, H Gupta, W Wang, C L Thompson, M M Knight.
       OBJECTIVE: Cartilage health is maintained in response to a range of mechanical stimuli including compressive, shear and tensile strains and associated alterations in osmolality. The osmotic-sensitive ion channel Transient Receptor Potential Vanilloid 4 (TRPV4) is required for mechanotransduction. Mechanical stimuli inhibit interleukin-1β (IL-1β) mediated inflammatory signalling, however the mechanism is unclear. This study aims to clarify the role of TRPV4 in this response.
    DESIGN: TRPV4 activity was modulated glycogen synthase kinase (GSK205 antagonist or GSK1016790 A (GSK101) agonist) in articular chondrocytes and cartilage explants in the presence or absence of IL-1β, mechanical (10% cyclic tensile strain (CTS), 0.33 Hz, 24hrs) or osmotic loading (200mOsm, 24hrs). Nitric oxide (NO), prostaglandin E2 (PGE2) and sulphated glycosaminoglycan (sGAG) release and cartilage biomechanics were analysed. Alterations in post-translational tubulin modifications and primary cilia length regulation were examined.
    RESULTS: In isolated chondrocytes, mechanical loading inhibited IL-1β mediated NO and PGE2 release. This response was inhibited by GSK205. Similarly, osmotic loading was anti-inflammatory in cells and explants, this response was abrogated by TRPV4 inhibition. In explants, GSK101 inhibited IL-1β mediated NO release and prevented cartilage degradation and loss of mechanical properties. Upon activation, TRPV4 cilia localisation was increased resulting in histone deacetylase 6 (HDAC6)-dependent modulation of soluble tubulin and altered cilia length regulation.
    CONCLUSION: Mechanical, osmotic or pharmaceutical activation of TRPV4 regulates HDAC6-dependent modulation of ciliary tubulin and is anti-inflammatory. This study reveals for the first time, the potential of TRPV4 manipulation as a novel therapeutic mechanism to supress pro-inflammatory signalling and cartilage degradation.
    Keywords:  Cartilage; Cilia; Hypo-osmolarity; IL-1β; Mechanotransduction; TRPV4
    DOI:  https://doi.org/10.1016/j.joca.2020.08.002
  8. Hum Genet. 2021 Jan 03.
    Bi-allelic BRWD1 variants cause male infertility with asthenoteratozoospermia and likely primary ciliary dyskinesia.
    Ting Guo, Chao-Feng Tu, Dan-Hui Yang, Shui-Zi Ding, Cheng Lei, Rong-Chun Wang, Lv Liu, Xi Kang, Xiao-Qing Shen, Yi-Feng Yang, Zhi-Ping Tan, Yue-Qiu Tan, Hong Luo.
      Genetics-associated asthenoteratozoospermia is often seen in patients with multiple morphological abnormalities of the sperm flagella (MMAF). Although 24 causative genes have been identified, these explain only approximately half of patients with MMAF. Since sperm flagella and motile cilia (especially respiratory cilia) have similar axonemal structures, many patients with MMAF also exhibit respiratory symptoms, such as recurrent airway infection, chronic sinusitis, and bronchiectasis, which are frequently associated with primary ciliary dyskinesia (PCD), another recessive disorder. Here, exome sequencing was conducted to evaluate the genetic cause in 53 patients with MMAF and classic PCD/PCD-like symptoms. Two homozygous missense variants and a compound-heterozygous variant in the BRWD1 gene were identified in three unrelated individuals. BRWD1 staining was detected in the whole flagella and respiratory cilia of normal controls but was absent in BRWD1-mutated individuals. Transmission electron microscopy and immunostaining demonstrated that BRWD1 deficiency in human affected respiratory cilia and sperm flagella differently, as the absence of outer and inner dynein arms in sperm flagellum and respiratory cilia, while with a decreased number and outer doublet microtubule defects of respiratory cilia. To our knowledge, this is the first report of a BRWD1-variant-related disease in humans, manifesting as an autosomal recessive form of MMAF and PCD/PCD-like symptoms. Our data provide a basis for further exploring the molecular mechanism of BRWD1 gene during spermatogenesis and ciliogenesis.
    DOI:  https://doi.org/10.1007/s00439-020-02241-4
  9. Front Cell Dev Biol. 2020 ;8 581340
    Host Antiviral Response Suppresses Ciliogenesis and Motile Ciliary Functions in the Nasal Epithelium.
    Qianmin Chen, Kai Sen Tan, Jing Liu, Hsiao Hui Ong, Suizi Zhou, Hongming Huang, Hailing Chen, Yew Kwang Ong, Mark Thong, Vincent T Chow, Qianhui Qiu, De-Yun Wang.
       Background: Respiratory viral infections are one of the main drivers of development and exacerbation for chronic airway inflammatory diseases. Increased viral susceptibility and impaired mucociliary clearance are often associated with chronic airway inflammatory diseases and served as risk factors of exacerbations. However, the links between viral susceptibility, viral clearance, and impaired mucociliary functions are unclear. Therefore, the objective of this study is to provide the insights into the effects of improper clearance of respiratory viruses from the epithelium following infection, and their resulting persistent activation of antiviral response, on mucociliary functions.
    Methods: In order to investigate the effects of persistent antiviral responses triggered by viral components from improper clearance on cilia formation and function, we established an in vitro air-liquid interface (ALI) culture of human nasal epithelial cells (hNECs) and used Poly(I:C) as a surrogate of viral components to simulate their effects toward re-epithelization and mucociliary functions of the nasal epithelium following damages from a viral infection.
    Results: Through previous and current viral infection expression data, we found that respiratory viral infection of hNECs downregulated motile cilia gene expression. We then further tested the effects of antiviral response activation on the differentiation of hNECs using Poly(I:C) stimulation on differentiating human nasal epithelial stem/progenitor cells (hNESPCs). Using this model, we observed reduced ciliated cell differentiation compared to goblet cells, reduced protein and mRNA in ciliogenesis-associated markers, and increased mis-assembly and mis-localization of ciliary protein DNAH5 following treatment with 25 μg/ml Poly(I:C) in differentiating hNECs. Additionally, the cilia length and ciliary beat frequency (CBF) were also decreased, which suggest impairment of ciliary function as well.
    Conclusion: Our results suggest that the impairments of ciliogenesis and ciliary function in hNECs may be triggered by specific expression of host antiviral response genes during re-epithelization of the nasal epithelium following viral infection. This event may in turn drive the development and exacerbation of chronic airway inflammatory diseases.
    Keywords:  antiviral response; cilia; human nasal epithelial cells (hNECs); mucociliary barrier; respiratory virus
    DOI:  https://doi.org/10.3389/fcell.2020.581340
  10. Biochem Biophys Res Commun. 2021 Jan 05. pii: S0006-291X(20)32270-1. [Epub ahead of print]539 28-33
    Functional inhibition of fatty acid binding protein 4 ameliorates impaired ciliogenesis in GCs.
    Yooju Jung, Sung Min Cho, Sungsoo Kim, Jae-Ho Cheong, Ho Jeong Kwon.
      Ciliogenesis is often impaired in some cancer cells, leading to acceleration of cancer phenotypes such as cell migration and proliferation. From the investigation of primary cilia of 16 gastric cancer cells (GCs), we found that GCs could be grouped into four primary cilia (PC)-positive GCs and 12 PC-negative GCs. The proliferation of the PC-positive GCs was lower than that of PC-negative GCs. To explore the role of fatty acid binding protein 4 (FABP4), which is a known oncogenic factor, in ciliogenesis, FABP4 expression and function were inhibited by transfection of cells with short interfering RNA targeting FABP4 (siFABP4) or FABP4 inhibitor treatment. Notably, the proliferation and migration of the cilia-forming GCs was effectively suppressed by inhibition of FABP4. In addition, the primary cilia in GCs were restored by a factor greater than two, suggesting a negative role of FABP4 in ciliogenesis in these GCs and FABP4 as a potential anticancer target.
    Keywords:  FABP4; Gastric cancer; Primary cilia; Small molecule
    DOI:  https://doi.org/10.1016/j.bbrc.2020.12.083
  11. PLoS Biol. 2021 Jan 06. 19(1): e3001060
    Ectoderm to mesoderm transition by down-regulation of actomyosin contractility.
    Leily Kashkooli, David Rozema, Lina Espejo-Ramirez, Paul Lasko, François Fagotto.
      Collective migration of cohesive tissues is a fundamental process in morphogenesis and is particularly well illustrated during gastrulation by the rapid and massive internalization of the mesoderm, which contrasts with the much more modest movements of the ectoderm. In the Xenopus embryo, the differences in morphogenetic capabilities of ectoderm and mesoderm can be connected to the intrinsic motility of individual cells, very low for ectoderm, highly for mesoderm. Surprisingly, we find that these seemingly deep differences can be accounted for simply by differences in Rho-kinases (Rock)-dependent actomyosin contractility. We show that Rock inhibition is sufficient to rapidly unleash motility in the ectoderm and confer it with mesoderm-like properties. In the mesoderm, this motility is dependent on 2 negative regulators of RhoA, the small GTPase Rnd1 and the RhoGAP Shirin/Dlc2/ArhGAP37. Both are absolutely essential for gastrulation. At the cellular and tissue level, the 2 regulators show overlapping yet distinct functions. They both contribute to decrease cortical tension and confer motility, but Shirin tends to increase tissue fluidity and stimulate dispersion, while Rnd1 tends to favor more compact collective migration. Thus, each is able to contribute to a specific property of the migratory behavior of the mesoderm. We propose that the "ectoderm to mesoderm transition" is a prototypic case of collective migration driven by a down-regulation of cellular tension, without the need for the complex changes traditionally associated with the epithelial-to-mesenchymal transition.
    DOI:  https://doi.org/10.1371/journal.pbio.3001060
  12. Redox Biol. 2020 Dec 25. pii: S2213-2317(20)31047-8. [Epub ahead of print]40 101842
    RhoA GTPase phosphorylated at tyrosine 42 by src kinase binds to β-catenin and contributes transcriptional regulation of vimentin upon Wnt3A.
    Jae-Gyu Kim, Shohel Mahmud, Jung Ki Min, Yoon-Beom Lee, Hyunbin Kim, Dong-Chul Kang, Hwee-Seon Park, Jihye Seong, Jae-Bong Park.
      In the Wnt canonical pathway, Wnt3A has been known to stabilize β-catenin. In the non-canonical Wnt signaling pathway, Wnt is known to activate Rho GTPases. The correlation between canonical and non-canonical pathways by Wnt signaling, however, has not been well elucidated. Here, we identified that Wnt3A promoted superoxide generation, leading to Tyr42 phosphorylation of RhoA through activations of c-Src and Rho-dependent coiled coil kinase 2 (ROCK2) and phosphorylation of p47phox, a component of NADPH oxidase. Wnt3A also induced accumulation of β-catenin along with activations of RhoA and ROCK1. Concurrently, ROCK1 was able to phosphorylate GSK-3β at Ser9, which phosphorylated Src at Ser51 and Ser492 residues, leading to Src inactivation through dephosphorylation of Tyr416 during the late period of Wnt3A treatment. Meanwhile, p-Tyr42 RhoA bound to β-catenin via the N-terminal domain of β-catenin, thereby leading to the nuclear translocation of p-Tyr42 RhoA/β-catenin complex. Notably, p-Tyr42 RhoA as well as β-catenin was associated with the promoter of Vim, leading to increased expression of vimentin. In addition, stomach cancer patients harboring higher expressed p-Tyr42 Rho levels revealed the much poorer survival probability. Therefore, we propose that p-Tyr42 RhoA is crucial for transcriptional regulation of specific target genes in the nucleus by binding to their promoters and involved in tumorigenesis.
    Keywords:  Cancer; GSK-3β; Src; Superoxide; Wnt3A; p-Tyr42 RhoA; β-Catenin
    DOI:  https://doi.org/10.1016/j.redox.2020.101842
  13. Theranostics. 2021 ;11(2): 614-648
    Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing.
    Song Zhang, Jiangtao Shen, Dali Li, Yiyun Cheng.
      CRISPR/Cas9 genome editing has gained rapidly increasing attentions in recent years, however, the translation of this biotechnology into therapy has been hindered by efficient delivery of CRISPR/Cas9 materials into target cells. Direct delivery of CRISPR/Cas9 system as a ribonucleoprotein (RNP) complex consisting of Cas9 protein and single guide RNA (sgRNA) has emerged as a powerful and widespread method for genome editing due to its advantages of transient genome editing and reduced off-target effects. In this review, we summarized the current Cas9 RNP delivery systems including physical approaches and synthetic carriers. The mechanisms and beneficial roles of these strategies in intracellular Cas9 RNP delivery were reviewed. Examples in the development of stimuli-responsive and targeted carriers for RNP delivery are highlighted. Finally, the challenges of current Cas9 RNP delivery systems and perspectives in rational design of next generation materials for this promising field will be discussed.
    Keywords:  CRISPR; RNP; genome editing; nanoparticles; polymers
    DOI:  https://doi.org/10.7150/thno.47007
  14. EMBO Rep. 2021 Jan 07. e51127
    Gradual centriole maturation associates with the mitotic surveillance pathway in mouse development.
    Cally Xiao, Marta Grzonka, Charlotte Meyer-Gerards, Miriam Mack, Rebecca Figge, Hisham Bazzi.
      Centrosomes, composed of two centrioles and pericentriolar material, organize mitotic spindles during cell division and template cilia during interphase. The first few divisions during mouse development occur without centrioles, which form around embryonic day (E) 3. However, disruption of centriole biogenesis in Sas-4 null mice leads to embryonic arrest around E9. Centriole loss in Sas-4-/- embryos causes prolonged mitosis and p53-dependent cell death. Studies in vitro discovered a similar USP28-, 53BP1-, and p53-dependent mitotic surveillance pathway that leads to cell cycle arrest. In this study, we show that an analogous pathway is conserved in vivo where 53BP1 and USP28 are upstream of p53 in Sas-4-/- embryos. The data indicate that the pathway is established around E7 of development, four days after the centrioles appear. Our data suggest that the newly formed centrioles gradually mature to participate in mitosis and cilia formation around the beginning of gastrulation, coinciding with the activation of mitotic surveillance pathway upon centriole loss.
    Keywords:  53BP1; SAS-4; USP28; centrosomes; p53
    DOI:  https://doi.org/10.15252/embr.202051127
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