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https://www.readbyqxmd.com/read/30045985/evasion-strategy-of-human-cytomegalovirus-to-escape-interferon-%C3%AE-induced-apobec3g-editing-activity
#1
Sara Pautasso, Ganna Galitska, Valentina Dell'Oste, Matteo Biolatti, Rachele Cagliani, Diego Forni, Marco De Andrea, Marisa Gariglio, Manuela Sironi, Santo Landolfo
The apolipoprotein B editing enzyme catalytic subunit 3 (APOBEC3) is a family of DNA cytosine deaminases that mutate and inactivate viral genomes by single-strand DNA editing, thus providing an innate immune response against a wide range of DNA and RNA viruses. In particular, APOBEC3A (A3A), a member of the APOBEC3 family, is induced by human cytomegalovirus (HCMV) in decidual tissues where it efficiently restricts HCMV replication, thereby acting as an intrinsic innate immune effector at the maternal-fetal interface...
July 25, 2018: Journal of Virology
https://www.readbyqxmd.com/read/30043294/model-legumes-functional-genomics-tools-in-medicago-truncatula
#2
Luis A Cañas, José Pío Beltrán
Many researchers have sought along the last two decades a legume species that could serve as a model system for genetic studies to resolve specific developmental or metabolic processes that cannot be studied in other model plants. Nitrogen fixation, nodulation, compound leaf, inflorescence and plant architecture, floral development, pod formation, secondary metabolite biosynthesis, and other developmental and metabolic aspects are legume-specific or show important differences with those described in Arabidopsis thaliana, the most studied model plant...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/30038938/efficient-enrichment-of-gene-modified-primary-t-cells-via-ccr5-targeted-integration-of-mutant-dihydrofolate-reductase
#3
Biswajit Paul, Guillermo S Romano Ibarra, Nicholas Hubbard, Teresa Einhaus, Alexander Astrakhan, David J Rawlings, Hans-Peter Kiem, Christopher W Peterson
Targeted gene therapy strategies utilizing homology-driven repair (HDR) allow for greater control over transgene integration site, copy number, and expression-significant advantages over traditional vector-mediated gene therapy with random genome integration. However, the relatively low efficiency of HDR-based strategies limits their clinical application. Here, we used HDR to knock in a mutant dihydrofolate reductase (mDHFR) selection gene at the gene-edited CCR5 locus in primary human CD4+ T cells and selected for mDHFR-modified cells in the presence of methotrexate (MTX)...
June 15, 2018: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/30026135/current-strategies-in-the-non-clinical-safety-assessment-of-biologics-new-targets-new-molecules-new-challenges
#4
Frank R Brennan, Laura Andrews, Antonio R Arulanandam, Jorg Blumel, Jim Fikes, Christine Grimaldi, Janice Lansita, Lise I Loberg, Tim MacLachlan, Mark Milton, Suezanne Parker, Jay Tibbitts, Jayanthi Wolf, Krishna P Allamneni
Nonclinical safety testing of biopharmaceuticals can present significant challenges to human risk assessment with these innovative and often complex drugs. Emerging topics in this field were discussed recently at the 2016 Annual US BioSafe General Membership meeting. The presentations and subsequent discussions from the main sessions are summarized. The topics covered included: (i) specialty biologics (oncolytic virus, gene therapy, and gene editing-based technologies), (ii) the value of non-human primates (NHPs) for safety assessment, (iii) challenges in the safety assessment of immuno-oncology drugs (T cell-dependent bispecifics, checkpoint inhibitors, and costimulatory agonists), (iv) emerging therapeutic approaches and modalities focused on microbiome, oligonucleotide, messenger ribonucleic acid (mRNA) therapeutics, (v) first in human (FIH) dose selection and the minimum anticipated biological effect level (MABEL), (vi) an update on current regulatory guidelines, International Council for Harmonization (ICH) S1, S3a, S5, S9 and S11 and (vii) breakout sessions that focused on bioanalytical and PK/PD challenges with bispecific antibodies, cytokine release in nonclinical studies, determining adversity and NOAEL for biologics, the value of second species for toxicology assessment and what to do if there is no relevant toxicology species...
July 17, 2018: Regulatory Toxicology and Pharmacology: RTP
https://www.readbyqxmd.com/read/30021904/enhancer-control-of-mir-155-expression-in-epstein-barr-virus-infected-b-cells
#5
C David Wood, Thomas Carvell, Andrea Gunnell, Opeoluwa O Ojeniyi, Cameron Osborne, Michelle J West
The oncogenic microRNA miR-155 is the most frequently upregulated miRNA in Epstein-Barr virus (EBV)-positive B cell malignancies and is upregulated in other non-viral lymphomas. Both the EBV nuclear antigen 2 (EBNA2), and B cell transcription factor, interferon regulatory factor 4 (IRF4) are known to activate transcription of the host cell gene from which miR-155 is processed ( miR-155HG , BIC). EBNA2 also activates IRF4 transcription indicating that EBV may upregulate miR-155 through direct and indirect mechanisms...
July 18, 2018: Journal of Virology
https://www.readbyqxmd.com/read/30019807/simultaneous-crispr-cas9-mediated-editing-of-cassava-eif4e-isoforms-ncbp-1-and-ncbp-2-reduces-cassava-brown-streak-disease-symptom-severity-and-incidence
#6
Michael A Gomez, Z Daniel Lin, Theodore Moll, Raj Deepika Chauhan, Luke Hayden, Kelley Renninger, Getu Beyene, Nigel J Taylor, J Carrington, B Staskawicz, R Bart
Cassava brown streak disease (CBSD) is a major constraint on cassava yields in East and Central Africa and threatens production in West Africa. CBSD is caused by two species of positive sense RNA viruses belonging to the family Potyviridae, genus Ipomovirus: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). Diseases caused by the family Potyviridae require the interaction of viral genome-linked protein (VPg) and host eukaryotic translation initiation factor 4E (eIF4E) isoforms...
July 18, 2018: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/30018062/stem-cell-derived-clade-f-aavs-mediate-high-efficiency-homologous-recombination-based-genome-editing
#7
Laura J Smith, Jason Wright, Gabriella Clark, Taihra Ul-Hasan, Xiangyang Jin, Abigail Fong, Manasa Chandra, Thia St Martin, Hillard Rubin, David Knowlton, Jeff L Ellsworth, Yuman Fong, Kamehameha K Wong, Saswati Chatterjee
The precise correction of genetic mutations at the nucleotide level is an attractive permanent therapeutic strategy for human disease. However, despite significant progress, challenges to efficient and accurate genome editing persist. Here, we report a genome editing platform based upon a class of hematopoietic stem cell (HSC)-derived clade F adeno-associated virus (AAV), which does not require prior nuclease-mediated DNA breaks and functions exclusively through BRCA2-dependent homologous recombination. Genome editing is guided by complementary homology arms and is highly accurate and seamless, with no evidence of on-target mutations, including insertion/deletions or inclusion of AAV inverted terminal repeats...
July 31, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29989433/oral-mucosa-derived-induced-pluripotent-stem-cells-from-patients-with-ectrodactyly-ectodermal-dysplasia-clefting-syndrome
#8
Marta Trevisan, Gualtiero Alvisi, Vanessa Barbaro, Luisa Barzon, Paolo Raffa, Angelo Migliorati, Giovanna Desole, Silvia Ruzittu, Giulia Masi, Enzo Di Iorio, Giorgio Palù
Ectrodactyly-Ectodermal dysplasia-Clefting (EEC) syndrome is a rare monogenic disease with autosomal dominant inheritance caused by mutations in the TP63 gene, leading to progressive corneal keratinocyte loss, limbal stem cell deficiency (LSCD), and eventually blindness. Currently, there is no treatment available to cure or slow down the keratinocyte loss. Human oral mucosal epithelial stem cells (hOMESCs), which are a mixed population of keratinocyte precursor stem cells, are used as source of autologous tissue for treatment of bilateral LSCD...
July 10, 2018: Cellular Reprogramming
https://www.readbyqxmd.com/read/29981111/an-introduction-to-plant-tissue-culture-advances-and-perspectives
#9
Victor M Loyola-Vargas, Neftalí Ochoa-Alejo
Plant tissue culture techniques are the most frequently used biotechnological tools for basic and applied purposes ranging from investigation on plant developmental processes, functional gene studies, commercial plant micropropagation, generation of transgenic plants with specific industrial and agronomical traits, plant breeding and crop improvement, virus elimination from infected materials to render high-quality healthy plant material, preservation and conservation of germplasm of vegetative propagated plant crops, and rescue of threatened or endangered plant species...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29961753/crispr-cas9-mediated-deletion-of-mir-146a-enhances-antiviral-response-in-hiv-1-infected-cells
#10
Yan Teng, Mingqi Luo, Ting Yu, Lang Chen, Qiuling Huang, Shuliang Chen, Linlin Xie, Yan Zeng, Fan Luo, Hairong Xiong, Yuanyuan Liu, Wei Hou, Yong Feng
The human immunodeficiency virus type 1 (HIV-1) causes persistent infection in human and induces miR-146a expression in infected cells. miR-146a represses the innate immune response by inhibiting the expression of TRAF6 and IRAK1 genes, thus negatively controls the NF-κB-related cytokines and interferon stimulated genes. Here we reported that lentiviral CRISPR/Cas9 system was highly efficient in introducing mutations in the precursor miR-146a genomic sequences, resulting in a loss of miR-146a expression and function...
July 2, 2018: Genes and Immunity
https://www.readbyqxmd.com/read/29957233/genome-editing-of-induced-pluripotent-stem-cells-to-decipher-cardiac-channelopathy-variant
#11
Priyanka Garg, Angelos Oikonomopoulos, Haodong Chen, Yingxin Li, Chi Keung Lam, Karim Sallam, Marco Perez, Robert L Lux, Michael C Sanguinetti, Joseph C Wu
BACKGROUND: The long QT syndrome (LQTS) is an arrhythmogenic disorder of QT interval prolongation that predisposes patients to life-threatening ventricular arrhythmias such as Torsades de pointes and sudden cardiac death. Clinical genetic testing has emerged as the standard of care to identify genetic variants in patients suspected of having LQTS. However, these results are often confounded by the discovery of variants of uncertain significance (VUS), for which there is insufficient evidence of pathogenicity...
July 3, 2018: Journal of the American College of Cardiology
https://www.readbyqxmd.com/read/29929184/viral-delivery-of-genome-modifying-proteins-for-cellular-reprogramming
#12
REVIEW
Jacob Giehm Mikkelsen
Following the successful development of virus-based gene vehicles for genetic therapies, exploitation of viruses as carriers of genetic tools for cellular reprogramming and genome editing should be right up the street. However, whereas persistent, potentially life-long gene expression is the main goal of conventional genetic therapies, tools and bits for genome engineering should ideally be short-lived and active only for a limited time. Although viral vector systems have already been adapted for potent genome editing both in vitro and in vivo, regulatable gene expression systems or self-limiting expression circuits need to be implemented limiting exposure of chromatin to genome-modifying enzymes...
June 18, 2018: Current Opinion in Genetics & Development
https://www.readbyqxmd.com/read/29925651/pigs-lacking-the-scavenger-receptor-cysteine-rich-domain-5-of-cd163-are-resistant-to-prrsv-1-infection
#13
Christine Burkard, Tanja Opriessnig, Alan J Mileham, Tomasz Stadejek, Tahar Ait-Ali, Simon G Lillico, C Bruce A Whitelaw, Alan L Archibald
Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) has a narrow host cell tropism, limited to cells of the monocyte/macrophage lineage. CD163 protein is expressed at high levels on the surface of specific macrophage types and a soluble form is circulating in blood. CD163 has been described as a fusion receptor for PRRSV, with the scavenger receptor cysteine-rich domain 5 (SRCR5) region having been shown to be the interaction site for the virus.As reported earlier, we have generated pigs in which Exon 7 of the CD163 gene has been deleted using CRISPR/Cas9 editing in pig zygotes...
June 20, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29921752/phage-genetic-engineering-using-crispr%C3%A2-cas-systems
#14
REVIEW
Asma Hatoum-Aslan
Since their discovery over a decade ago, the class of prokaryotic immune systems known as CRISPR⁻Cas have afforded a suite of genetic tools that have revolutionized research in model organisms spanning all domains of life. CRISPR-mediated tools have also emerged for the natural targets of CRISPR⁻Cas immunity, the viruses that specifically infect bacteria, or phages. Despite their status as the most abundant biological entities on the planet, the majority of phage genes have unassigned functions. This reality underscores the need for robust genetic tools to study them...
June 19, 2018: Viruses
https://www.readbyqxmd.com/read/29906476/adar1-affects-hcv-infection-by-modulating-innate-immune-response
#15
Maria Pujantell, Sandra Franco, Iván Galván-Femenía, Roger Badia, Marc Castellví, Edurne Garcia-Vidal, Bonaventura Clotet, Rafael de Cid, Cristina Tural, Miguel A Martínez, Eva Riveira-Muñoz, José A Esté, Ester Ballana
The hepatitis C virus (HCV) is a globally prevalent infectious pathogen. As many as 80% of people infected with HCV do not control the virus and develop a chronic infection. Response to interferon (IFN) therapy is widely variable in chronic HCV infected patients, suggesting that HCV has evolved mechanisms to suppress and evade innate immunity responsible for its control and elimination. Adenosine deaminase acting on RNA 1 (ADAR1) is a relevant factor in the regulation of the innate immune response. The loss of ADAR1 RNA-editing activity and the resulting loss of inosine bases in RNA are critical in producing aberrant RLR-mediated innate immune response, mediated by RNA sensors MDA5 and RIG-I...
August 2018: Antiviral Research
https://www.readbyqxmd.com/read/29903474/crispr-cas13a-prospects-for-plant-virus-resistance
#16
Muhammad Zuhaib Khan, Imran Amin, Amir Hameed, Shahid Mansoor
CRISPR-Cas13a is an efficient RNA targeting and editing tool characterized recently in prokaryotes. This system can be recruited to engineer resistance against plant RNA viruses and regulate gene expression. We discuss some important achievements in using the CRISPR-Cas13a system to confer resistance against plant RNA viruses.
June 11, 2018: Trends in Biotechnology
https://www.readbyqxmd.com/read/29899171/novel-approaches-for-the-treatment-of-familial-hypercholesterolemia-current-status-and-future-challenges
#17
Long Jiang, Lu-Ya Wang, Xiao-Shu Cheng
Familial hypercholesterolemia (FH) is an autosomal-dominant disorder that is characterized by high plasma low-density lipoprotein cholesterol (LDL-c) levels and an increased risk of cardiovascular disease. Despite the use of high-dose statins and the recent addition of proprotein convertase subtilisin/kexin type 9 inhibitors as a treatment option, many patients with homozygous FH fail to achieve optimal reductions of LDL-c levels. Gene therapy has become one of the most promising research directions for contemporary life sciences and is a potential treatment option for FH...
June 13, 2018: Journal of Atherosclerosis and Thrombosis
https://www.readbyqxmd.com/read/29872154/hiv-1-inhibition-in-cells-with-cxcr4-mutant-genome-created-by-crispr-cas9-and-piggybac-recombinant-technologies
#18
Shuai Liu, Qiankun Wang, Xiao Yu, Yilin Li, Yandan Guo, Zhepeng Liu, Fuyun Sun, Wei Hou, Chunmei Li, Li Wu, Deyin Guo, Shuliang Chen
The C-X-C chemokine receptor type 4 (CXCR4) is one of the major co-receptors for human immunodeficiency virus type 1 (HIV-1) entry and is considered an important therapeutic target. However, its function in maintaining the development of hematopoietic stem cells (HSC) makes it difficult to be used for HIV-1 gene therapy with HSC transplantation. A previous report showed that the natural CXCR4 P191A mutant inhibits HIV-1 infection without any defect in HSC differentiation, which could provide a basis for the development of new approaches for HIV-1 gene therapy...
June 5, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29857485/absence-of-a3z3-related-hypermutations-in-the-env-and-vif-proviral-genes-in-fiv-naturally-infected-cats
#19
Lucía Cano-Ortiz, Dennis Maletich Junqueira, Juliana Comerlato, André Zani, Cristina Santos Costa, Paulo Michel Roehe, Ana Cláudia Franco
Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3; A3) proteins comprise an important family of restriction factors that produce hypermutations on proviral DNA and are able to limit virus replication. Vif, an accessory protein present in almost all lentiviruses, counteracts the antiviral A3 activity. Seven haplotypes of APOBEC3Z3 ( A3Z3 ) were described in domestic cats (hap I⁻VII), and in-vitro studies have demonstrated that these proteins reduce infectivity of vif-defective feline immunodeficiency virus (FIV)...
May 31, 2018: Viruses
https://www.readbyqxmd.com/read/29847792/site-specific-gene-editing-of-human-hematopoietic-stem-cells-for-x-linked-hyper-igm-syndrome
#20
Caroline Y Kuo, Joseph D Long, Beatriz Campo-Fernandez, Satiro de Oliveira, Aaron R Cooper, Zulema Romero, Megan D Hoban, Alok V Joglekar, Georgia R Lill, Michael L Kaufman, Sorel Fitz-Gibbon, Xiaoyan Wang, Roger P Hollis, Donald B Kohn
X-linked hyper-immunoglobulin M (hyper-IgM) syndrome (XHIM) is a primary immunodeficiency due to mutations in CD40 ligand that affect immunoglobulin class-switch recombination and somatic hypermutation. The disease is amenable to gene therapy using retroviral vectors, but dysregulated gene expression results in abnormal lymphoproliferation in mouse models, highlighting the need for alternative strategies. Here, we demonstrate the ability of both the transcription activator-like effector nuclease (TALEN) and clustered regularly interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) platforms to efficiently drive integration of a normal copy of the CD40L cDNA delivered by Adeno-Associated Virus...
May 29, 2018: Cell Reports
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