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https://www.readbyqxmd.com/read/29222170/imprecision-and-dna-break-repair-biased-towards-incompatible-end-joining-in-leukemia
#1
Franz Josef Gassner, Maria Schubert, Stefan Rebhandl, Karina Spandl, Nadja Zaborsky, Kemal Catakovic, Stephanie Blaimer, Daniel Hebenstreit, Richard Greil, Roland Geisberger
Cancer is a genetic disease caused by mutations and chromosomal abnormalities which contribute to uncontrolled cell growth. In addition, cancer cells can rapidly respond to conventional and targeted therapies by accumulating novel and often specific genetic lesions leading to acquired drug resistance and relapsing disease. In chronic lymphocytic leukemia (CLL), however, diverse chromosomal aberrations often occur. In many cases, improper repair of DNA double strand breaks (DSBs) is a major source for genomic abnormalities...
December 8, 2017: Molecular Cancer Research: MCR
https://www.readbyqxmd.com/read/29216270/generation-and-characterization-of-kctd15-mutations-in-zebrafish
#2
Alison Heffer, Gregory D Marquart, Allisan Aquilina-Beck, Nabil Saleem, Harold A Burgess, Igor B Dawid
Potassium channel tetramerization domain containing 15 (Kctd15) was previously found to have a role in early neural crest (NC) patterning, specifically delimiting the region where NC markers are expressed via repression of transcription factor AP-2a and inhibition of Wnt signaling. We used transcription activator-like effector nucleases (TALENs) to generate null mutations in zebrafish kctd15a and kctd15b paralogs to study the in vivo role of Kctd15. We found that while deletions producing frame-shift mutations in each paralog showed no apparent phenotype, kctd15a/b double mutant zebrafish are smaller in size and show several phenotypes including some affecting the NC, such as expansion of the early NC domain, increased pigmentation, and craniofacial defects...
2017: PloS One
https://www.readbyqxmd.com/read/29208373/roles-of-maternal-wnt8a-transcripts-in-axis-formation-in-zebrafish
#3
Hiromu Hino, Akiko Nakanishi, Ryoko Seki, Tsubasa Aoki, Etsuro Yamaha, Atsuo Kawahara, Takashi Shimizu, Masahiko Hibi
In early zebrafish development, the program for dorsal axis formation begins soon after fertilization. Previous studies suggested that dorsal determinants (DDs) localize to the vegetal pole, and are transported to the dorsal blastomeres in a microtubule-dependent manner. The DDs activate the canonical Wnt pathway and induce dorsal-specific genes that are required for dorsal axis formation. Among wnt-family genes, only the wnt8a mRNA is reported to localize to the vegetal pole in oocytes and to induce the dorsal axis, suggesting that Wnt8a is a candidate DD...
December 2, 2017: Developmental Biology
https://www.readbyqxmd.com/read/29196142/new-transgenic-models-of-parkinson-s-disease-using-genome-editing-technology
#4
J A Cota-Coronado, S Sandoval-Ávila, Y P Gaytan-Dávila, N F Diaz, B Vega-Ruiz, E Padilla-Camberos, N E Díaz-Martínez
INTRODUCTION: Parkinson's disease (PD) is the second most common neurodegenerative disorder. It is characterised by selective loss of dopaminergic neurons in the substantia nigra pars compacta, which results in dopamine depletion, leading to a number of motor and non-motor symptoms. DEVELOPMENT: In recent years, the development of new animal models using nuclease-based genome-editing technology (ZFN, TALEN, and CRISPR/Cas9 nucleases) has enabled the introduction of custom-made modifications into the genome to replicate key features of PD, leading to significant advances in our understanding of the pathophysiology of the disease...
November 28, 2017: Neurología: Publicación Oficial de la Sociedad Española de Neurología
https://www.readbyqxmd.com/read/29186020/applications-of-alternative-nucleases-in-the-age-of-crispr-cas9
#5
REVIEW
Tuhin K Guha, David R Edgell
Breakthroughs in the development of programmable site-specific nucleases, including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), meganucleases (MNs), and most recently, the clustered regularly interspaced short palindromic repeats (CRISPR) associated proteins (including Cas9) have greatly enabled and accelerated genome editing. By targeting double-strand breaks to user-defined locations, the rates of DNA repair events are greatly enhanced relative to un-catalyzed events at the same sites...
November 29, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29179035/generation-of-a-talen-mediated-p63-knock-in-in-human-induced-pluripotent-stem-cells
#6
Yuki Kobayashi, Ryuhei Hayashi, Andrew J Quantock, Kohji Nishida
The expression of p63 in surface ectodermal cells during development of the cornea, skin, oral mucosa and olfactory placodes is integral to the process of cellular self-renewal and the maintenance of the epithelial stem cell status. Here, we used TALEN technology to generate a p63 knock-in (KI) human induced pluripotent stem (hiPS) cell line in which p63 expression can be visualized via enhanced green fluorescent protein (EGFP) expression. The KI-hiPS cells maintained pluripotency and expressed the stem cell marker gene, ΔNp63α...
October 31, 2017: Stem Cell Research
https://www.readbyqxmd.com/read/29163669/current-status-and-perspectives-of-genome-editing-technology-for-microalgae
#7
REVIEW
Seungjib Jeon, Jong-Min Lim, Hyung-Gwan Lee, Sung-Eun Shin, Nam Kyu Kang, Youn-Il Park, Hee-Mock Oh, Won-Joong Jeong, Byeong-Ryool Jeong, Yong Keun Chang
Genome editing techniques are critical for manipulating genes not only to investigate their functions in biology but also to improve traits for genetic engineering in biotechnology. Genome editing has been greatly facilitated by engineered nucleases, dubbed molecular scissors, including zinc-finger nuclease (ZFN), TAL effector endonuclease (TALEN) and clustered regularly interspaced palindromic sequences (CRISPR)/Cas9. In particular, CRISPR/Cas9 has revolutionized genome editing fields with its simplicity, efficiency and accuracy compared to previous nucleases...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/29150002/gene-editing-and-crispr-therapeutics-strategies-taught-by-cell-and-gene-therapy
#8
Juan C Ramirez
A few years ago, we assisted in the demonstration for the first time of the revolutionary idea of a type of adaptive-immune system in the bacteria kingdom. This system, named CRISPR, and variants engineered in the lab, have been demonstrated as functional with extremely high frequency and fidelity in almost all eukaryotic cells studied to date. The capabilities of this RNA-guided nuclease have added to the interest that was announced with the advent of previous technologies for genome editing tools, such as ZFN and TALEN...
2017: Progress in Molecular Biology and Translational Science
https://www.readbyqxmd.com/read/29138463/talen-mediated-shift-of-mitochondrial-dna-heteroplasmy-in-melas-ipscs-with-m-13513g-a-mutation
#9
Naoki Yahata, Yuji Matsumoto, Minoru Omi, Naoki Yamamoto, Ryuji Hata
Induced pluripotent stem cells (iPSCs) are suitable for studying mitochondrial diseases caused by mitochondrial DNA (mtDNA) mutations. Here, we generated iPSCs from a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with the m.13513G>A mutation. The patient's dermal fibroblasts were reprogrammed, and we established two iPSC clones with and without mutant mtDNA. Furthermore, we tried to decrease mutant mtDNA level in iPSCs using transcription activator-like effector nucleases (TALENs)...
November 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29130159/the-future-of-crispr-applications-in-the-lab-the-clinic-and-society
#10
Soren H Hough, Ayokunmi Ajetunmobi
CRISPR (clustered regularly interspaced short palindromic repeats) has emerged as one of the premiere biological tools of the century. Even more so than older genome editing techniques such as TALENs and ZFNs, CRISPR provides speed and ease-of-use heretofore unheard of in agriculture, the environment and human health. The ability to map the function of virtually every component of the genome in a scalable, multiplexed manner is unprecedented. Once those regions have been explored, CRISPR also presents an opportunity to take advantage of endogenous cellular repair pathways to change and precisely edit the genome [1-3]...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/29124278/targeted-gene-editing-in-human-pluripotent-stem-cells-using-site-specific-nucleases
#11
Sylvia Merkert, Ulrich Martin
Introduction of induced pluripotent stem cell (iPSC) technology and site-directed nucleases brought a major breakthrough in the development of regenerative therapies and biomedical research. With the advancement of ZFNs, TALENs, and the CRISPR/Cas9 technology, straightforward and precise manipulation of the genome of human pluripotent stem cells (PSC) became possible, allowing relatively easy and fast generation of gene knockouts, integration of transgenes, or even introduction of single nucleotide changes for correction or introduction of disease-specific mutations...
November 10, 2017: Advances in Biochemical Engineering/biotechnology
https://www.readbyqxmd.com/read/29123113/talen-mediated-functional-correction-of-human-ipsc-derived-macrophages-in-context-of-hereditary-pulmonary-alveolar-proteinosis
#12
Alexandra Kuhn, Mania Ackermann, Claudio Mussolino, Toni Cathomen, Nico Lachmann, Thomas Moritz
Hereditary pulmonary alveolar proteinosis (herPAP) constitutes a rare, life threatening lung disease characterized by the inability of alveolar macrophages to clear the alveolar airspaces from surfactant phospholipids. On a molecular level, the disorder is defined by a defect in the CSF2RA gene coding for the GM-CSF receptor alpha-chain (CD116). As therapeutic options are limited, we currently pursue a cell and gene therapy approach aiming for the intrapulmonary transplantation of gene-corrected macrophages derived from herPAP-specific induced pluripotent stem cells (herPAP-iPSC) employing transcriptional activator-like effector nucleases (TALENs)...
November 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29120617/live-visualization-of-hiv-1-proviral-dna-using-a-dual-color-labeled-crispr-system
#13
Yingxin Ma, Mingxiu Wang, Wei Li, Zhi-Ping Zhang, Xiaowei Zhang, Tianwei Tan, Zongqiang Cui, Xian-En Zhang
HIV latency is one of the major problems in HIV/AIDS cure. Imaging single-copy integrated proviral HIV DNA in host cell has both virology and clinical significance but remains technical challenge. Here, we developed a dual-color labeled CRISPR system to image the HIV-1 integrated proviral DNA in latently infected cells. The pair of CRISPRs was fluorescently labeled with two different color QDs using two alternative bioorthogonal ligation reactions. Integrated HIV-sequences are successfully mapped based on the co-localized signals of QDs in living cells...
November 9, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/29115572/talens-mediated-homozygous-ccr5%C3%AE-32-mutations-endow-cd4-u87-cells-with-resistance-against-hiv%C3%A2-1-infection
#14
Ai Qing Yu, Yan Ding, Zhi Yong Lu, Yan Zhe Hao, Zhi Ping Teng, Shi Rong Yan, Dong Sheng Li, Yi Zeng
Since evidence suggests that transplantation of bone marrow stem cells with the C‑C chemokine receptor type 5 (CCR5)Δ32/Δ32 genotype may cure patients infected with human immunodeficiency virus (HIV)‑1, the present study aimed to reproduce the CCR5Δ32 mutation in cluster of differentiation (CD)4+ U87 cells using genome engineering methods. A modified transcription activator‑like effector nucleases (TALENs) technique, combined with homologous recombination for site‑specific, size‑controlled and homozygous DNA deletions, was used to reproduce the homozygous CCR5Δ32 mutation in CD4+ U87 cells...
October 26, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29110389/equivalence-testing-for-similarity-in-bioassays-using-bioequivalence-criteria-on-the-relative-bioactivity
#15
Corine Baljé-Volkers, Thembile Mzolo, Erik Talens, Pieta IJzerman-Boon, Edwin Van den Heuvel
Similarity in bioassays means that the test preparation behaves as a dilution of the standard preparation with respect to their biological effect. Thus, similarity must be investigated to confirm this biological property. Historically, this was typically conducted with traditional hypothesis testing, but this has received substantial criticism. Failing to reject similarity does not imply that the 2 preparations are similar. Also, rejecting similarity when bioassay variability is small might simply demonstrate a nonrelevant deviation in similarity...
November 6, 2017: Pharmaceutical Statistics
https://www.readbyqxmd.com/read/29100547/efficient-generation-of-p53-biallelic-knockout-diannan-miniature-pigs-via-talens-and-somatic-cell-nuclear-transfer
#16
Youfeng Shen, Kaixiang Xu, Zaimei Yuan, Jianxiong Guo, Heng Zhao, Xuezeng Zhang, Lu Zhao, Yubo Qing, Honghui Li, Weirong Pan, Baoyu Jia, Hong-Ye Zhao, Hong-Jiang Wei
BACKGROUND: Pigs have many features that make them attractive as biomedical models for various diseases, including cancer. P53 is an important tumor suppressor gene that exerts a central role in protecting cells from oncogenic transformation and is mutated in a large number of human cancers. P53 mutations occur in almost every type of tumor and in over 50% of all tumors. In a recent publication, pigs with a mutated P53 gene were generated that resulted in lymphoma and renal and osteogenic tumors...
November 3, 2017: Journal of Translational Medicine
https://www.readbyqxmd.com/read/29100435/effect-of-talen-mediated-il-6-knockout-on-cell-proliferation-apoptosis-invasion-and-anti-cancer-therapy-in-hepatocellular-carcinoma-hcc-lm3-cells
#17
Peng-Yuan Zhuang, Ke-Wei Zhang, Jian-Dong Wang, Xue-Ping Zhou, Ying-Bin Liu, Zhi-Wei Quan, Jun Shen
Purpose: To determine the exact effect of Interleukin-6 (IL-6) on tumor cell proliferation, apoptosis, invasion, and anti-cancer therapy in hepatocellular carcinoma (HCC). Experimental Design: IL-6 was disrupted by transcription activator-like effector nucleases (TALEN) in HCCLM3 cells, and was used to evaluate the role of IL-6 on tumor cell proliferation, apoptosis, invasion and key signaling pathways involved in sorafenib and/or IFNα therapy. Results: IL-6 has no direct effect on cell proliferation and invasion but promotes cell apoptosis and up-regulate IL-33 and VEGF-A expression...
September 29, 2017: Oncotarget
https://www.readbyqxmd.com/read/29097757/mapping-the-expression-of-the-sex-determining-factor-doublesex1-in-daphnia-magna-using-a-knock-in-reporter
#18
Quang Dang Nong, Nur Syafiqah Mohamad Ishak, Tomoaki Matsuura, Yasuhiko Kato, Hajime Watanabe
Sexually dimorphic traits are common and widespread among animals. The expression of the Doublesex-/Mab-3-domain (DM-domain) gene family has been widely studied in model organisms and has been proven to be essential for the development and maintenance of sex-specific traits. However, little is known about the detailed expression patterns in non-model organisms. In the present study, we demonstrated the spatiotemporal expression of the DM-domain gene, doublesex1 (dsx1), in the crustacean Daphnia magna, which parthenogenetically produces males in response to environmental cues...
November 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29094286/genome-editing-in-livestock-are-we-ready-for-a-revolution-in-animal-breeding-industry
#19
REVIEW
Jinxue Ruan, Jie Xu, Ruby Yanru Chen-Tsai, Kui Li
Genome editing is a powerful technology that can efficiently alter the genome of organisms to achieve targeted modification of endogenous genes and targeted integration of exogenous genes. Current genome-editing tools mainly include ZFN, TALEN and CRISPR/Cas9, which have been successfully applied to all species tested including zebrafish, humans, mice, rats, monkeys, pigs, cattle, sheep, goats and others. The application of genome editing has quickly swept through the entire biomedical field, including livestock breeding...
November 1, 2017: Transgenic Research
https://www.readbyqxmd.com/read/29082723/-genome-editing-focus-on-the-off-target-effects
#20
Xiubin He, Feng Gu
Breakthroughs of genome-editing in recent years have paved the way to develop new therapeutic strategies. These genome-editing tools mainly include Zinc-finger nucleases (ZFNs), Transcription activator-like effector nucleases (TALENs), and clustered regulatory interspaced short palindromic repeat (CRISPR)/Cas-based RNA-guided DNA endonucleases. However, off-target effects are still the major issue in genome editing, and limit the application in gene therapy. Here, we summarized the cause and compared different detection methods of off-targets...
October 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
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