Chuanbo Sun, Bing Li, Miaomiao Yang, Ruirui Guo, Simin Yuan, Jichang Wang, Hao Hu
GPAM (glycerol-3-phosphateacyltransferase1) is a mitochondrial enzyme that catalyze an essential step in glycerolphospholipids and triacylglycerol biosynthesis process. Loss-of-function mutation of GPAM has been shown to lead to hypomyelination of corticospinal tract in cerebral palsy patient. To model this rare disease with human brain organoid, we generated a GPAM knockout human embryonic stem cell line SYSUe-008-A by CRISPR/cas9. The GPAM knockout cell line maintains a normal karyotype and shows comparable level of pluripotent stem cell marker expression and differentiation potential as wild-type human embryonic stem cells...
March 19, 2021: Stem Cell Research
Amelia D Wallace, Thomas A Sasani, Jordan Swanier, Brooke L Gates, Jeff Greenland, Brent S Pedersen, Katherine E Varley, Aaron R Quinlan
A substantial fraction of the human genome is difficult to interrogate with short-read DNA sequencing technologies due to paralogy, complex haplotype structures, or tandem repeats. Long-read sequencing technologies, such as Oxford Nanopore's MinION, enable direct measurement of complex loci without introducing many of the biases inherent to short-read methods, though they suffer from relatively lower throughput. This limitation has motivated recent efforts to develop amplification-free strategies to target and enrich loci of interest for subsequent sequencing with long reads...
2021: PloS One
Ana K Ibarrola-Vannucci, Luis M De Pablos, Lissette Retana-Moreira, Alberto Cornet-Gómez, Teresa Cruz-Bustos, Alejandro G Schijman, José L Ramírez, Susana Vílchez, Antonio Osuna
BACKGROUND: Chagas disease is the third most important neglected tropical disease. There is no vaccine available, and only two drugs are generally prescribed for the treatment, both of which with a wide range of side effects. Our study of T. cruzi PHBs revealed a pleiotropic function in different stages of the parasite, participating actively in the transformation of the non-infective replicative epimastigote form into metacyclic trypomastigotes and also in the multiplication of intracellular amastigotes...
April 8, 2021: PLoS Neglected Tropical Diseases
Yu Zhang, Qian Wang, Jing Wang, Xinjing Tang
The CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR-associated protein 9) is a most powerful tool and has been widely used in gene editing and gene regulation since its discovery. However, wild-type CRISPR-Cas9 suffers from off-target effects and low editing efficiency. To overcome these limitations, engineered Cas9 proteins have been extensively investigated. In addition to Cas9 protein engineering, chemically synthesized guide RNAs have been developed to improve the efficiency and specificity of genome editing as well as spatiotemporal controllability, which broadens the biological applications of CRISPR-Cas9 gene editing system and increases their potentials as therapeutics...
March 18, 2021: ChemPlusChem
Emrah Gumusgoz, Dikran R Guisso, Sahba Kasiri, Jun Wu, Matthew Dear, Brandy Verhalen, Silvia Nitschke, Sharmistha Mitra, Felix Nitschke, Berge A Minassian
Many adult and most childhood neurological diseases have a genetic basis. CRISPR/Cas9 biotechnology holds great promise in neurological therapy, pending the clearance of major delivery, efficiency, and specificity hurdles. We applied CRISPR/Cas9 genome editing in its simplest modality, namely inducing gene sequence disruption, to one adult and one pediatric disease. Adult polyglucosan body disease is a neurodegenerative disease resembling amyotrophic lateral sclerosis. Lafora disease is a severe late childhood onset progressive myoclonus epilepsy...
April 8, 2021: Neurotherapeutics: the Journal of the American Society for Experimental NeuroTherapeutics
Vasanth Thamodaran, Sonam Rani, Shaji R Velayudhan
Induced pluripotent stem cells (iPSCs) generated from patients are a valuable tool for disease modelling, drug screening, and studying the functions of cell/tissue-specific genes. However, for this research, isogenic iPSC lines are important for comparison of phenotypes in the wild type and mutant differentiated cells generated from the iPSCs. The advent of gene editing technologies to correct or generate mutations helps in the generation of isogenic iPSC lines with the same genetic background. Due to the ease of programming, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas9-based gene editing tools have gained pace in gene manipulation studies, including investigating complex diseases like cancer...
April 9, 2021: Methods in Molecular Biology
Abozar Ghorbani, Shima Hadifar, Roya Salari, Keramatollah Izadpanah, Michal Burmistrz, Alireza Afsharifar, Mohammad Hadi Eskandari, Ali Niazi, Christopher E Denes, G Gregory Neely
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) together with CRISPR-associated (Cas) proteins have catalysed a revolution in genetic engineering. Native CRISPR-Cas systems exist in many bacteria and archaea where they provide an adaptive immune response through sequence-specific degradation of an invading pathogen's genome. This system has been reconfigured for use in genome editing, drug development, gene expression regulation, diagnostics, the prevention and treatment of cancers, and the treatment of genetic and infectious diseases...
April 8, 2021: Transgenic Research
Laetitia Maroc, Cécile Fairhead
This short paper aims to review what our recent studies in the Nakaseomyces yeasts, principally Candida glabrata, reveal about the evolution of the mating-type switching system and its components, as well as about the repair of chromosomal double-strand breaks in this clade. In the model yeast Saccharomyces cerevisiae, the study of mating-type switching has, over the years, led to major discoveries in how cells process chromosomal breaks. Indeed, in this species, switching, which allows every haploid cell to produce cells of opposite mating types that can mate together, is initiated by the Ho endonuclease, linking sexual reproduction to a programmed chromosomal cut...
April 8, 2021: Current Genetics
Yanyun Li, Shuting Liao, Pengying Mei, Yueyun Pan, Yu Zhang, Xiangzi Zheng, Yakun Xie, Ying Miao
Cross talking between natural senescence and cell death in response to pathogen attack is an interesting topic; however, its action mechanism is kept open. In this study, 33 OsWRKY genes were obtained by screening with leaf aging procedure through RNA-seq dataset, and 11 of them were confirmed a significant altered expression level in the flag leaves during aging by using the reverse transcript quantitative PCR (RT-qPCR). Among them, the OsWRKY2 , OsWRKY14 , OsWRKY26 , OsWRKY69 , and OsWRKY93 members exhibited short-term alteration in transcriptional levels in response to Magnaporthe grisea infection...
2021: Frontiers in Plant Science
Alissa Hendricks-Wenger, Kenneth N Aycock, Margaret A Nagai-Singer, Sheryl Coutermarsh-Ott, Melvin F Lorenzo, Jessica Gannon, Kyungjun Uh, Kayla Farrell, Natalie Beitel-White, Rebecca M Brock, Alexander Simon, Holly A Morrison, Joanne Tuohy, Sherrie Clark-Deener, Eli Vlaisavljevich, Rafael V Davalos, Kiho Lee, Irving C Allen
New therapies to treat pancreatic cancer are direly needed. However, efficacious interventions lack a strong preclinical model that can recapitulate patients' anatomy and physiology. Likewise, the availability of human primary malignant tissue for ex vivo studies is limited. These are significant limitations in the biomedical device field. We have developed RAG2/IL2RG deficient pigs using CRISPR/Cas9 as a large animal model with the novel application of cancer xenograft studies of human pancreatic adenocarcinoma...
April 7, 2021: Scientific Reports
Guannan Zhao, Wenjing Zhang, Peixin Dong, Hidemichi Watari, Yuqi Guo, Lawrence M Pfeffer, Gabor Tigyi, Junming Yue
BACKGROUND: Epithelial to mesenchymal transition (EMT) contributes to tumor metastasis and chemoresistance. Eukaryotic initiation factor 5A2 (EIF5A2) is highly expressed in a variety of human cancers but rarely expressed in normal tissues. While EIF5A2 has oncogenic activity in several cancers and contributes to tumor metastasis, its role in ovarian cancer is unknown. In this study, we investigate whether EIF5A2 contributes to ovarian tumor metastasis by promoting EMT. METHODS: To investigate the role of EIF5A2, we knocked out (KO) EIF5A2 using lentiviral CRISPR/Cas9 nickase in high invasive SKOV3 and OVCAR8 cells and overexpressed EIF5A2 in low invasive OVCAR3 cells using lentiviral vector...
April 7, 2021: Cell & Bioscience
Channabasavaiah B Gurumurthy, Aidan R O'Brien, Rolen M Quadros, John Adams, Pilar Alcaide, Shinya Ayabe, Johnathan Ballard, Surinder K Batra, Marie-Claude Beauchamp, Kathleen A Becker, Guillaume Bernas, David Brough, Francisco Carrillo-Salinas, Wesley Chan, Hanying Chen, Ruby Dawson, Victoria DeMambro, Jinke D'Hont, Katharine Dibb, James D Eudy, Lin Gan, Jing Gao, Amy Gonzales, Anyonya Guntur, Huiping Guo, Donald W Harms, Anne Harrington, Kathryn E Hentges, Neil Humphreys, Shiho Imai, Hideshi Ishii, Mizuho Iwama, Eric Jonasch, Michelle Karolak, Bernard Keavney, Nay-Chi Khin, Masamitsu Konno, Yuko Kotani, Yayoi Kunihiro, Imayavaramban Lakshmanan, Catherine Larochelle, Catherine B Lawrence, Lin Li, Volkhard Lindner, Xian-De Liu, Gloria Lopez-Castejon, Andrew Loudon, Jenna Lowe, Loydie Jerome-Majeweska, Taiji Matsusaka, Hiromi Miura, Yoshiki Miyasaka, Benjamin Morpurgo, Katherine Motyl, Yo-Ichi Nabeshima, Koji Nakade, Toshiaki Nakashiba, Kenichi Nakashima, Yuichi Obata, Sanae Ogiwara, Mariette Ouellet, Leif Oxburgh, Sandra Piltz, Ilka Pinz, Moorthy P Ponnusamy, David Ray, Ronald J Redder, Clifford J Rosen, Nikki Ross, Mark T Ruhe, Larisa Ryzhova, Ane M Salvador, Sabrina Shameen Alam, Radislav Sedlacek, Karan Sharma, Chad Smith, Katrien Staes, Lora Starrs, Fumihiro Sugiyama, Satoru Takahashi, Tomohiro Tanaka, Andrew Trafford, Yoshihiro Uno, Leen Vanhoutte, Frederique Vanrockeghem, Brandon J Willis, Christian S Wright, Yuko Yamauchi, Xin Yi, Kazuto Yoshimi, Xuesong Zhang, Yu Zhang, Masato Ohtsuka, Satyabrata Das, Daniel J Garry, Tino Hochepied, Paul Thomas, Jan Parker-Thornburg, Antony D Adamson, Atsushi Yoshiki, Jean-Francois Schmouth, Andrei Golovko, William R Thompson, K C Kent Lloyd, Joshua A Wood, Mitra Cowan, Tomoji Mashimo, Seiya Mizuno, Hao Zhu, Petr Kasparek, Lucy Liaw, Joseph M Miano, Gaetan Burgio
No abstract text is available yet for this article.
April 7, 2021: Genome Biology
Hui Yang, Haoyi Wang, Rudolf Jaenisch
No abstract text is available yet for this article.
April 7, 2021: Genome Biology
Kathryn Wright, Kumudika de Silva, Karren M Plain, Auriol C Purdie, Tamika A Blair, Iain G Duggin, Warwick J Britton, Stefan H Oehlers
Pathogenic mycobacteria actively dysregulate protective host immune signalling pathways during infection to drive the formation of permissive granuloma microenvironments. Dynamic regulation of host microRNA (miRNA) expression is a conserved feature of mycobacterial infections across host-pathogen pairings. Here we examine the role of miR-206 in the zebrafish model of Mycobacterium marinum infection, which allows investigation of the early stages of granuloma formation. We find miR-206 is upregulated following infection by pathogenic M...
April 7, 2021: PLoS Pathogens
Rui Li, Runling Zhang, Ping Tan, Meng Wang, Yuqing Chen, Jiawei Zhang, Dongsheng Han, Yanxi Han, Jinming Li, Rui Zhang
BACKGROUND: Mismatch repair deficiency (dMMR) status induced by MLH1 protein deficiency plays a pivotal role in therapeutic decision-making for cancer patients. Appropriate quality control (QC) materials are necessary for monitoring the accuracy of MLH1 protein deficiency assays used in clinical laboratories. METHODS: CRISPR/Cas9 technology was used to edit the MLH1 gene of GM12878Cas9 cells to establish MLH1 protein-deficient cell lines. The positive cell lines were screened and validated by Sanger sequencing, Western blot (WB), and next-generation sequencing (NGS) and were then used to prepare formalin-fixed, paraffin-embedded (FFPE) samples through xenografting...
April 7, 2021: Journal of Clinical Laboratory Analysis
Haruo Nakano, Shiori Kawai, Yusaku Ooki, Tomoki Chiba, Chiharu Ishii, Takumi Nozawa, Hisako Utsuki, Mariko Umemura, Shigeru Takahashi, Yuji Takahashi
Activating transcription factor 5 (ATF5) is a stress-responsive transcription factor that belongs to the cAMP response element-binding protein (CREB)/ATF family, and is essential for the differentiation and survival of sensory neurons in murine olfactory organs. However, the study of associated proteins and target genes for ATF5 has been hampered due to the limited availability of immunoprecipitation-grade ATF5 antibodies. To overcome this issue, we generated hemagglutinin (HA)-tag knock-in mice for ATF5 using CRISPR/Cas9-mediated genome editing with one-step electroporation in oviducts (i-GONAD)...
April 7, 2021: Cell and Tissue Research
Daisuke Matsumoto, Wataru Nomura
Genome editing technology commenced in 1996 with the discovery of the first zinc-finger nuclease. Application of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) associated protein 9 (Cas9) technology to genome editing of mammalian cells allowed researchers to use genome editing more easily and cost-effectively. However, one of the technological problems that remains to be solved is "off-target effects", which are unexpected mutations in nontarget DNA. One significant improvement in genome editing technology has been achieved with molecular/protein engineering...
April 7, 2021: Bioconjugate Chemistry
Tsugumi Ueda, Tomoaki Iwayama, Kiwako Tomita, Shuji Matsumoto, Mizuho Iwashita, Phan Bhongsatiern, Hiromi Sakashita, Chiharu Fujihara, Masahide Takedachi, Shinya Murakami
Periodontal ligament (PDL) possesses a stem/progenitor population to maintain the homeostasis of periodontal tissue. However, transcription factors that regulate this population have not yet been identified. Thus, we aimed to identify a molecule related to the osteogenic differentiation of PDL progenitors using a single cell-based strategy in this study. We first devised a new protocol to isolate PDL cells from the surface of adult murine molars and established 35 new single cell-derived clones from the PDL explant...
April 6, 2021: Scientific Reports
Ying-Chen Xia, Jian-Hua Zha, Yong-Hua Sang, Hui Yin, Guo-Qiu Xu, Jie Zhen, Yan Zhang, Ben-Tong Yu
Activation of adenosine monophosphate-activated protein kinase (AMPK) is able to produce significant anti-non-small cell lung cancer (NSCLC) cell activity. ASP4132 is an orally active and highly effective AMPK activator. The current study tested its activity against NSCLC cells. In primary NSCLC cells and established cell lines (A549 and NCI-H1944) ASP4132 potently inhibited cell growth, proliferation and cell cycle progression as well as cell migration and invasion. Robust apoptosis activation was detected in ASP4132-treated NSCLC cells...
April 6, 2021: Cell Death & Disease
Raquel S Negreiros, Noelia Lander, Miguel A Chiurillo, Anibal E Vercesi, Roberto Docampo
Pyruvate is the final metabolite of glycolysis and can be converted into acetyl coenzyme A (acetyl-CoA) in mitochondria, where it is used as the substrate for the tricarboxylic acid cycle. Pyruvate availability in mitochondria depends on its active transport through the heterocomplex formed by the mitochondrial pyruvate carriers 1 and 2 (MPC1/MPC2). We report here studies on MPC1/MPC2 of Trypanosoma cruzi , the etiologic agent of Chagas disease. Endogenous tagging of T. cruzi MPC1 ( TcMPC1 ) and TcMPC2 with 3× c-Myc showed that both encoded proteins colocalize with MitoTracker to the mitochondria of epimastigotes...
April 6, 2021: MBio
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