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https://www.readbyqxmd.com/read/30517854/shortening-the-half-life-of-cas9-maintains-its-gene-editing-ability-and-reduces-neuronal-toxicity
#1
Su Yang, Shihua Li, Xiao-Jiang Li
Virus-mediated expression of CRISPR/Cas9 is commonly used for genome editing in animal brains to model or treat neurological diseases, but the potential neurotoxicity of overexpressing bacterial Cas9 in the mammalian brain remains unknown. Through RNA sequencing (RNA-seq) analysis, we find that virus-mediated expression of Cas9 influences the expression of genes involved in neuronal functions. Reducing the half-life of Cas9 by tagging with geminin, whose expression is regulated by the cell cycle, maintains the genome editing capacity of Cas9 but significantly alleviates neurotoxicity...
December 4, 2018: Cell Reports
https://www.readbyqxmd.com/read/30517747/allele-specific-genome-editing-using-crispr-cas9-is-associated-with-loss-of-heterozygosity-in-diploid-yeast
#2
Arthur R Gorter de Vries, Lucas G F Couwenberg, Marcel van den Broek, Pilar de la Torre Cortés, Jolanda Ter Horst, Jack T Pronk, Jean-Marc G Daran
Targeted DNA double-strand breaks (DSBs) with CRISPR-Cas9 have revolutionized genetic modification by enabling efficient genome editing in a broad range of eukaryotic systems. Accurate gene editing is possible with near-perfect efficiency in haploid or (predominantly) homozygous genomes. However, genomes exhibiting polyploidy and/or high degrees of heterozygosity are less amenable to genetic modification. Here, we report an up to 99-fold lower gene editing efficiency when editing individual heterozygous loci in the yeast genome...
December 5, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/30517736/extensive-crispr-rna-modification-reveals-chemical-compatibility-and-structure-activity-relationships-for-cas9-biochemical-activity
#3
Daniel O'Reilly, Zachary J Kartje, Eman A Ageely, Elise Malek-Adamian, Maryam Habibian, Annabelle Schofield, Christopher L Barkau, Kushal J Rohilla, Lauren B DeRossett, Austin T Weigle, Masad J Damha, Keith T Gagnon
CRISPR (clustered regularly interspaced short palindromic repeat) endonucleases are at the forefront of biotechnology, synthetic biology and gene editing. Methods for controlling enzyme properties promise to improve existing applications and enable new technologies. CRISPR enzymes rely on RNA cofactors to guide catalysis. Therefore, chemical modification of the guide RNA can be used to characterize structure-activity relationships within CRISPR ribonucleoprotein (RNP) enzymes and identify compatible chemistries for controlling activity...
December 4, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/30517136/essentiality-of-plasmodium-falciparum-plasmepsin-v
#4
Nonlawat Boonyalai, Christine R Collins, Fiona Hackett, Chrislaine Withers-Martinez, Michael J Blackman
The malaria parasite replicates within erythrocytes. The pathogenesis of clinical malaria is in large part due to the capacity of the parasite to remodel its host cell. To do this, intraerythrocytic stages of Plasmodium falciparum export more than 300 proteins that dramatically alter the morphology of the infected erythrocyte as well as its mechanical and adhesive properties. P. falciparum plasmepsin V (PfPMV) is an aspartic protease that processes proteins for export into the host erythrocyte and is thought to play a key role in parasite virulence and survival...
2018: PloS One
https://www.readbyqxmd.com/read/30516026/the-arrogance-of-teratology-a-brief-chronology-of-attitudes-throughout-history
#5
REVIEW
John M DeSesso
While the discipline of Teratology has existed for about 60 years, there has been a deep interest in the causes of human malformations for millennia. Absent the scientific method and acting on fervent beliefs that made sense to ancient/medieval populations, "mechanisms" were described and prognostications of future events were assigned to terata resulting in tragic (and unwarranted) sequelae. This article examines the collective beliefs and thinking within various eras in the hope of providing lessons to inform future behavior...
December 4, 2018: Birth Defects Research
https://www.readbyqxmd.com/read/30513454/overcoming-the-undesirable-crispr-cas9-expression-in-gene-correction
#6
Emily Xia, Rongqi Duan, Fushan Shi, Kyle E Seigel, Hartmut Grasemann, Jim Hu
The CRISPR-Cas9 system is attractive for gene therapy, as it allows for permanent genetic correction. However, as a new technology, Cas9 gene editing in clinical applications faces major challenges, such as safe delivery and gene targeting efficiency. Cas9 is a foreign protein to recipient cells; thus, its expression may prompt the immune system to eliminate gene-edited cells. To overcome these challenges, we have engineered a novel delivery system based on the helper-dependent adenoviral (HD-Ad) vector, which is capable of delivering genes to airway basal stem cells in vivo...
October 30, 2018: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/30513119/structural-basis-of-toxoplasma-gondii-perforin-like-protein-1-membrane-interaction-and-activity-during-egress
#7
Alfredo J Guerra, Ou Zhang, Constance M E Bahr, My-Hang Huynh, James DelProposto, William C Brown, Zdzislaw Wawrzak, Nicole M Koropatkin, Vern B Carruthers
Intracellular pathogens must egress from the host cell to continue their infectious cycle. Apicomplexans are a phylum of intracellular protozoans that have evolved members of the membrane attack complex and perforin (MACPF) family of pore forming proteins to disrupt cellular membranes for traversing cells during tissue migration or egress from a replicative vacuole following intracellular reproduction. Previous work showed that the apicomplexan Toxoplasma gondii secretes a perforin-like protein (TgPLP1) that contains a C-terminal Domain (CTD) which is necessary for efficient parasite egress...
December 4, 2018: PLoS Pathogens
https://www.readbyqxmd.com/read/30511992/crispr-craft-dna-editing-the-reconstructive-ladder
#8
Danny S Roh, Edward B-H Li, Eric C Liao
The clustered regularly interspaced short palindromic repeats (CRISPR) system of genome editing represents a major technological advance spanning all areas of genetics and downstream applications. CRISPR's potential impact on treating human disease encompasses all clinical specialties, including areas important to the plastic surgeon such as oncology, wound healing, immunology, and craniofacial malformations. Plastic surgeons should gain familiarity with this gene editing technology, and become active contributors and leaders in applying CRISPR to their respective areas of expertise...
November 2018: Plastic and Reconstructive Surgery
https://www.readbyqxmd.com/read/30510181/exocyst-dynamics-during-vesicle-tethering-and-fusion
#9
Syed Mukhtar Ahmed, Hisayo Nishida-Fukuda, Yuchong Li, W Hayes McDonald, Claudiu C Gradinaru, Ian G Macara
The exocyst is a conserved octameric complex that tethers exocytic vesicles to the plasma membrane prior to fusion. Exocyst assembly and delivery mechanisms remain unclear, especially in mammalian cells. Here we tagged multiple endogenous exocyst subunits with sfGFP or Halo using Cas9 gene-editing, to create single and double knock-in lines of mammary epithelial cells, and interrogated exocyst dynamics by high-speed imaging and correlation spectroscopy. We discovered that mammalian exocyst is comprised of tetrameric subcomplexes that can associate independently with vesicles and plasma membrane and are in dynamic equilibrium with octamer and monomers...
December 3, 2018: Nature Communications
https://www.readbyqxmd.com/read/30505938/functional-gene-knockout-of-nrf2-increases-chemosensitivity-of-human-lung-cancer-a549-cells-in-vitro-and-in-a-xenograft-mouse-model
#10
Pawel Bialk, Yichen Wang, Kelly Banas, Eric B Kmiec
Recent studies point to the evolution of drug resistance in lung cancer as being centered, at least in part, on the upregulation of various genes involved in controlling efflux or drug inactivation. Among the most important of these genes is Nuclear Factor Erythroid 2-Related Factor ( NRF2 ), considered the master regulator of 100-200 target genes involved in cellular responses to oxidative and/or electrophilic stress. With increased focus on the development of combinatorial approaches for cancer treatment, we utilized CRISPR/Cas9 to disable the NRF2 gene in lung cancer cells by disrupting the NRF2 nuclear export signal (NES) domain; phenotypically, the protein is largely blocked from transiting into the nucleus after translation...
December 21, 2018: Molecular Therapy Oncolytics
https://www.readbyqxmd.com/read/30504793/a-novel-cloning-strategy-for-one-step-assembly-of-multiplex-crispr-vectors
#11
Marc Zuckermann, Mario Hlevnjak, Haniyeh Yazdanparast, Marc Zapatka, David T W Jones, Peter Lichter, Jan Gronych
One key advantage of the CRISPR/Cas9 system in comparison with other gene editing approaches lies in its potential for multiplexing. Here, we describe an elaborate procedure that allows the assembly of multiple gRNA expression cassettes into a vector of choice within a single step, termed ASAP(Adaptable System for Assembly of multiplexed Plasmids)-cloning. We demonstrate the utility of ASAP-cloning for multiple CRISPR-mediated applications, including efficient multiplex gene editing, robust transcription activation and convenient analysis of Cas9 activity in the presence of multiple gRNAs...
November 30, 2018: Scientific Reports
https://www.readbyqxmd.com/read/30503558/triptonide-inhibits-human-nasopharyngeal-carcinoma-cell-growth-via-disrupting-lnc-rna-thor-igf2bp1-signaling
#12
Shu-Sheng Wang, Yan Lv, Xin-Chun Xu, Yun Zuo, Yu Song, Ge-Ping Wu, Pei-Hua Lu, Zhi-Qing Zhang, Min-Bin Chen
Advanced stage nasopharyngeal carcinoma (NPC) has a poor prognosis. Triptonide ("TN") is a small molecule monomer extract from the ancient Chinese herb Tripterygium wilfordii Hook. We show that TN, at nanomolar concentrations, potently inhibited survival and proliferation of multiple established and primary human NPC cells. TN induced NPC cell cycle arrest and apoptosis activation. NPC cell migration and invasion were also inhibited by TN. Importantly, TN was non-cytotoxic to nasopharyngeal epithelial cells...
November 29, 2018: Cancer Letters
https://www.readbyqxmd.com/read/30503261/%C3%AE-cell-replacement-after-gene-editing-of-a-neonatal-diabetes-causing-mutation-at-the-insulin-locus
#13
Shuangyu Ma, Ryan Viola, Lina Sui, Valentino Cherubini, Fabrizio Barbetti, Dieter Egli
Permanent neonatal diabetes mellitus (PNDM) can be caused by insulin mutations. We generated induced pluripotent stem cells from fibroblasts of a patient with PNDM and undetectable insulin at birth due to a homozygous mutation in the translation start site of the insulin gene. Differentiation of mutant cells resulted in insulin-negative endocrine stem cells expressing MAFA, NKX6.1, and chromogranin A. Correction of the mutation in stem cells and differentiation to pancreatic endocrine cells restored insulin production and insulin secretion to levels comparable to those of wild-type cells...
November 20, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/30501735/-crispr-cas9-system-and-its-advances-in-gene-therapy-of-hematologic-diseases-review
#14
Wan-Ying Li, Qing-Ping Gao, Hui Liu
In recent years, with the development of gene editing technology, the site-specific genome can be modified. The curability of genetic disease may be achieved by the use of gene editing techniques. As the simplicity, high specificity and economical efficiency, much attention has been paid to the CRISPR/Cas9 system, which was been widely used in research of molecular biology and other fields of life science. In this review, the mechanism for CR1SPR/Cas9 system and the progress of gene therapy, such as for hemophilia, betathalassaemia and chronic myeloid leukemia were summarized briefly...
December 2018: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/30499276/-interaction-of-bombyx-mori-aminopeptidase-n-and-cadherin-like-protein-with-bacillus-thuringiensis-cry1ac-toxin
#15
Ping Lin, Tingcai Cheng, Tieshan Feng, Jiao Gong, Chun Liu, Qingyou Xia
Bacillus thuringiensis (Bt) produces Cry toxins that are widely used as insecticides in agriculture and forestry. Receptors are important to elucidate the mode of interaction with Cry toxins and toxicity in lepidopteran insects. Here, we purified the Cry toxin from Bt and identified this toxin by flight mass spectrometry as Cry1Ac, and then recombinantly expressed aminopeptidase N (BmAPN6) and repeat domains of cadherin-like protein (CaLP) of B. mori. Using co-immunoprecipitation (co-IP), Far-Western blotting, and enzyme-linked immunosorbent assays (ELISAs), we identified the interaction between Cry1Ac and BmAPN6...
November 25, 2018: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/30498995/applications-of-pna-laden-nanoparticles-for-hematological-disorders
#16
REVIEW
Shipra Malik, Stanley Oyaghire, Raman Bahal
Safe and efficient genome editing has been an unmitigated goal for biomedical researchers since its inception. The most prevalent strategy for gene editing is the use of engineered nucleases that induce DNA damage and take advantage of cellular DNA repair machinery. This includes meganucleases, zinc-finger nucleases, transcription activator-like effector nucleases, and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas9) systems. However, the clinical viability of these nucleases is marred by their off-target cleavage activity (≥ 50% in RNA-guided endonucleases)...
November 29, 2018: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/30498505/strategies-for-engineering-reproductive-sterility-in-plantation-forests
#17
Steffi Fritsche, Amy L Klocko, Agnieszka Boron, Amy M Brunner, Glenn Thorlby
A considerable body of research exists concerning the development of technologies to engineer sterility in forest trees. The primary driver for this work has been to mitigate concerns arising from gene flow from commercial plantings of genetically engineered (GE) trees to non-GE plantations, or to wild or feral relatives. More recently, there has been interest in the use of sterility technologies as a means to mitigate the global environmental and socio-economic damage caused by the escape of non-native invasive tree species from planted forests...
2018: Frontiers in Plant Science
https://www.readbyqxmd.com/read/30498239/gene-therapy-for-lysosomal-storage-diseases-and-peroxisomal-diseases
#18
REVIEW
Toya Ohashi
Gene therapies for lysosomal storage diseases (LSD) and peroxisomal diseases (PD) are rapidly advancing. Most LSDs and PDs are characterized by brain involvement, prompting the development of therapies targeting the brain. There are two types of gene therapy for brain involvement in LSD and PD, i.e., the direct transfer of a therapeutic gene into brain cells and hematopoietic stem cell-targeted gene therapy. The rationale for the latter approach is that brain microglia are derived from hematopoietic cells. Thus, gene-corrected hematopoietic cells migrate into the brain and differentiate into microglial cells...
November 29, 2018: Journal of Human Genetics
https://www.readbyqxmd.com/read/30498087/a-positive-feedback-mechanism-ensures-proper-assembly-of-the-functional-inner-centromere-during-mitosis-in-human-cells
#19
Cai Liang, Zhenlei Zhang, Qinfu Chen, Haiyan Yan, Miao Zhang, Xingfeng Xiang, Qi Yi, Xuan Pan, Hankun Cheng, Fangwei Wang
The inner centromere region of a mitotic chromosome critically regulates sister chromatid cohesion and kinetochore-microtubule attachments. However, the molecular mechanism underlying inner centromere assembly remains elusive. Here, using CRISPR/Cas9-based gene editing in HeLa cells, we disrupted the interaction of Shugoshin 1 (Sgo1) with histone H2A phosphorylated on Thr-120 (H2ApT120) to selectively release Sgo1 from mitotic centromeres. Interestingly, cells expressing the H2ApT120-binding defective mutant of Sgo1 have an elevated rate of chromosome missegregation accompanied by weakened centromeric cohesion and decreased centromere accumulation of the chromosomal passenger complex (CPC), an integral part of the inner centromere and a key player in the correction of erroneous kinetochore-microtubule attachments...
November 29, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/30488815/-doing-crispr
#20
Dorothy Jane Dankel
Salmon farming is a key industry in Norway, with firsthand value of more than 60 billion Norwegian crowns in 2017. The salmon industry is a driving force for biotechnological applications in the marine sector. The recent release of the Atlantic salmon reference genome offers new opportunities to solve major aquaculture bottlenecks that currently limit expansion of the industry. One major bottleneck is the genetic impact of escaped farmed salmon on wild populations. To solve this problem, the industry can use sterile salmon in production...
2018: Politics and the Life Sciences: the Journal of the Association for Politics and the Life Sciences
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