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JOURNAL ARTICLE
Fu Guo, Yizi Huang, Peipei Qi, Guiwei Lian, Xingming Hu, Ning Han, Junhui Wang, Muyuan Zhu, Qian Qian, Hongwu Bian
Auxin regulates almost every aspect of plant growth and development and is perceived by the TIR1/AFB auxin co-receptor proteins differentially acting in concert with specific Aux/IAA transcriptional repressors. Little is known about the diverse functions of TIR1/AFB family members in species other than Arabidopsis. We created targeted OsTIR1 and OsAFB2-5 mutations in rice using CRISPR/Cas9 genome editing, and functionally characterized the roles of these five members in plant growth and development and auxinic herbicide resistance...
March 2021: New Phytologist
#22
JOURNAL ARTICLE
Hengji Zhan, Aolin Li, Zhiming Cai, Weiren Huang, Yuchen Liu
As a novel and robust gene-editing tool, the Clustered Regularly Interspaced Short Palindromic Repeats CRISPR-associated protein 9 (CRISPR-Cas9) system has revolutionized gene therapy. Plasmid vector delivery is the most commonly used method for integrating the CRISPR-Cas9 system into cells. However, such foreign cytosolic DNAs trigger an innate immune response (IIR) within cells, which can hinder gene editing by inhibiting transgene expression. Although some small molecules have been shown to avoid the action of IIR on plasmids, they only work on a single target and may also affect cell viability...
October 2020: Clinical and Translational Medicine
#23
REVIEW
Yaqi Li, Peiyuan Tang, Sanjun Cai, Junjie Peng, Guoqiang Hua
Three-dimensional cultured organoids have become a powerful in vitro research tool that preserves genetic, phenotypic and behavioral trait of in vivo organs, which can be established from both pluripotent stem cells and adult stem cells. Organoids derived from adult stem cells can be established directly from diseased epithelium and matched normal tissues, and organoids can also be genetically manipulated by CRISPR-Cas9 technology. Applications of organoids in basic research involve the modeling of human development and diseases, including genetic, infectious and malignant diseases...
November 2, 2020: Cell Regeneration
#24
JOURNAL ARTICLE
Yun Huang, Blerim Marovca, Susanne Dettwiler, Peter Karl Bode, Beat Bornhauser, Jean-Pierre Bourquin
No abstract text is available yet for this article.
October 2020: HemaSphere
#25
JOURNAL ARTICLE
Luiz Sergio Almeida Camargo, Joseph R Owen, Alison L Van Eenennaam, Pablo Juan Ross
Somatic cell nuclear transfer or cytoplasm microinjection have been used to generate genome-edited farm animals; however, these methods have several drawbacks that reduce their efficiency. This study aimed to develop electroporation conditions that allow delivery of CRISPR/Cas9 system to bovine zygotes for efficient gene knock-out. We optimized electroporation conditions to deliver Cas9:sgRNA ribonucleoproteins to bovine zygotes without compromising embryo development. Higher electroporation pulse voltage resulted in increased membrane permeability; however, voltages above 15 V/mm decreased embryo developmental potential...
2020: Frontiers in Genetics
#26
JOURNAL ARTICLE
Uyen Cao Chu, Sandeep Kumar, Amy Sigmund, Kari Johnson, Yinghong Li, Pamila Vongdeuane, Todd J Jones
Agrobacterium -mediated transformation of canola ( Brassica napus ) via hypocotyl segments has been a commonly used method for the past 30 years. While the hypocotyl-based method is well-established, it is not readily adapted to elite germplasm and the prolonged process is not ideal for a production transformation setting. We developed an Agrobacterium -mediated transformation method based on epicotyl and higher stem (internodal) segments that is efficient, rapid and amenable for high-throughput transformation and genome editing...
2020: Frontiers in Plant Science
#27
JOURNAL ARTICLE
JunXia Gao, LiZhe Liu, Chao Liu, ShuJuan Fan, LiRong Liu, ShuFeng Liu, Xiao-Hui Xian, Wen-Bin Li
Objective: Glutamate transporter-1 (GLT-1) and system x c - mediate glutamate uptake and release, respectively. Ceftriaxone has been reported to upregulate GLT-1 expression and improve cognitive decline in APP/PS1 mice. The aim of the present study was to elucidate the role of GLT-1 in ceftriaxone-mediated improvement on cognitive deficits and associated changes in xCT (catalytic subunit of system x c - ) expression and activity using GLT-1 knockdown APP/PS1 mice. Methods: GLT-1 knockdown (GLT-1± ) mice were generated in C57BL/6J mice using the CRISPR/Cas9 technique and crossed to APP/PS1 mice to generate GLT-1± APP/PS1 mice...
2020: Frontiers in Aging Neuroscience
#28
JOURNAL ARTICLE
Nicole A Hawkins, Sunita N Misra, Manuel Jurado, Seok Kyu Kang, Nicholas C Vierra, Kimberly Nguyen, Lisa Wren, Alfred L George, James S Trimmer, Jennifer A Kearney
Developmental and epileptic encephalopathies (DEE) are a group of severe epilepsies that usually present with intractable seizures, developmental delay, and often have elevated risk for premature mortality. Numerous genes have been identified as a monogenic cause of DEE, including KCNB1. The voltage-gated potassium channel KV 2.1, encoded by KCNB1, is primarily responsible for delayed rectifier potassium currents that are important regulators of excitability in electrically excitable cells, including neurons...
January 2021: Neurobiology of Disease
#29
JOURNAL ARTICLE
Dan Wang, Yongxin Guo, Huiming Li, Jiannan Li, Mingzi Ran, Juan Guo, Lu Yin, Shiyi Zhao, Qianzi Yang, Hailong Dong
BACKGROUND: The neuropeptide orexin promotes arousal from general anaesthesia, however the neuronal circuits that mediate this effect have not been defined. We investigated whether orexinergic neurones modulate the basal forebrain (BF) and locus coeruleus (LC) in emergence from anaesthesia. METHODS: Hcrtcre rats were generated using a CRISPR/Cas9-based approach. Viruses encoding optogenetic probes were injected into the perifornical lateral hypothalamic (PeFLH) area, optogenetic fibres were embedded in the PeFLH, BF, or LC, and changes in anaesthesia state under 1...
January 2021: British Journal of Anaesthesia
#30
JOURNAL ARTICLE
Yizhen Chen, Mingchuan Fu, Hao Li, Liguo Wang, Renzhong Liu, Zhanji Liu, Xianlong Zhang, Shuangxia Jin
Cotton (Gossypium hirsutum), the most important cash crop for natural textile fibers, meanwhile, represents the fifth largest source of vegetable oil for human consumption in the world. Typically, cottonseed oil contains three major fatty acids: 26% palmitic acid, 15% oleic acid, and 58% linoleic acid (Liu et al., 2002). The relatively high level of linoleic acid reduces oxidative stability of cottonseed oil, which can cause rancidity, a short shelf life and production of detrimental trans-fatty acids (Shockey et al...
November 1, 2020: Plant Biotechnology Journal
#31
JOURNAL ARTICLE
Qian Bu, Huaqin Zhang, Qian Liu, Yanping Dai, Qian Wei, Aiqin Xue, Yan Huang, Kai Zhong, Yina Huang, Hong Gao, Xiaobo Cen
N-acetylneuraminic acid synthase (NANS), the gene encoding the synthase for N-acetylneuraminic acid (NeuNAc; sialic acid), is closely associated with infantile-onset severe developmental delay and skeletal dysplasia. However, the role and the involved mechanisms of NANS functioning have not been fully understood to date. Here, we generated a homozygous NANS-knockout human induced pluripotent stem cell (iPSC) line, NCCSEDi001-A-1, via the CRISPR/Cas9-based gene editing method. The NCCSEDi001-A-1 cell line does not express NANS protein, but maintains a normal karyotype, pluripotency, and trilineage differentiation potential...
October 20, 2020: Stem Cell Research
#32
JOURNAL ARTICLE
Liang Huang, Zhan-Qi Dong, Fei-Fang Dong, Xi-Bo Yu, Zhi-Gang Hu, Na-Chuan Liao, Peng Chen, Cheng Lu, Min-Hui Pan
Apoptosis is a cellular defense mechanism used for the elimination of host cells infected by viruses. Viruses have evolved corresponding inhibitors of apoptosis genes to promote their replication. Anti-apoptosis-related genes, involved in baculovirus proliferation, have been proposed but it is unclear whether these genes can be manipulated in gene therapy. We constructed a transgenic silkworm, using the CRISPR/Cas9 system to knock out the BmNPV inhibitor of apoptosis 2 (iap2). The sequencing results showed that all the sequences could edit the target site of BmNPV iap2 gene...
October 29, 2020: International Journal of Biological Macromolecules
#33
JOURNAL ARTICLE
Jéssica Bassani Borges, Victor Fernandes de Oliveira, Glaucio Monteiro Ferreira, Bruna Los, Thais Kristini Almendros Afonso Barbosa, Elisangela da Silva Rodrigues Marçal, Carolina Dagli-Hernandez, Renata Caroline Costa de Freitas, Raul Hernandes Bortolin, Augusto Akira Mori, Thiago Dominguez Crespo Hirata, Helder Takashi Imoto Nakaya, Gisele Medeiros Bastos, Helena Strelow Thurow, Rodrigo Marques Gonçalves, Daniel Branco de Araujo, Henry Paulo Zatz, Adriana Bertolami, André Arpad Faludi, Marcelo Chiara Bertolami, Amanda Guerra de Moraes Rego Sousa, João Ítalo Dias França, Cinthia Elim Jannes, Alexandre da Costa Pereira, Marcelo Arruda Nakazone, Dorotéia Rossi Silva Souza, Tayanne Silva Carmo, Marcelo Ferraz Sampaio, Renata Gorjão, Tania Cristina Pithon-Curi, Patricia Moriel, Vivian Nogueira Silbiger, André Ducati Luchessi, Jéssica Nayara Góes de Araújo, Michel Satya Naslavsky, Jaqueline Yu Ting Wang, Thales Kronenberger, Alvaro Cerda, Hui Tzu Lin-Wang, Adriana Regina Garofalo, Cristina Moreno Fajardo, Rosario Dominguez Crespo Hirata, Mario Hiroyuki Hirata
BACKGROUND: Familial hypercholesterolemia (FH) is a genetic disease that affects millions of people worldwide. OBJECTIVES: The study protocol FHBGEP was design to investigate the main genomic, epigenomic, and pharmacogenomic factors associated with FH and polygenic hypercholesterolemia (PH). METHODS: FH patients will be enrolled at six research centers in Brazil. An exon-targeted gene strategy will be used to sequence a panel of 84 genes related to FH, PH, pharmacogenomics and coronary artery disease...
October 28, 2020: Research in Social & Administrative Pharmacy: RSAP
#34
JOURNAL ARTICLE
Alejandro Fulgencio-Covián, Mar Álvarez, Barry A Pepers, Arístides López-Márquez, Magdalena Ugarte, Belén Pérez, Willeke M C van Roon-Mom, Lourdes R Desviat, Eva Richard
Propionic acidemia (PA) is an inherited metabolic disease caused by mutations in the PCCA and PCCB genes. We have previously generated an induced pluripotent stem cell (iPSC) line (UAMi004-A) from a PA patient with the c.1218_1231del14ins12 (p.Gly407Argfs*14) homozygous mutation in the PCCB gene. Here, we report the generation of the isogenic control in which the mutation was genetically corrected using CRISPR/Cas9 technology. Off-target editing presence was excluded and the iPSCs had typical embryonic stem cell-like morphology and normal karyotype that expressed pluripotency markers and maintained their in vitro differentiation potential...
October 17, 2020: Stem Cell Research
#35
JOURNAL ARTICLE
Xiangcheng Tang, Zhigang Chen, Xuhua Tan, Lixia Luo, Xialin Liu, Lili Gong, David Wan-Cheng Li, Yizhi Liu
Mutations in the CRB1 gene reportedly cause early-onset autosomal recessive retinitis pigmentosa (RP), which can result in severe loss of vision at an early age. To investigate the mechanism of CRB1-knockout (CRB1-/- ) induced RP, we generated a subline of H9 human embryonic stem cells harboring frame shift mutations in a homozygous state in exon 2 of the CRB1 gene. This subline expressed pluripotent stem cell markers, presented a normal karyotype, and preserved the ability to differentiate into endodermal, mesodermal, and ectodermal lineages...
October 16, 2020: Stem Cell Research
#36
JOURNAL ARTICLE
Nobuaki Takahashi, Patricia Cho, Laura M Selfors, Hendrik J Kuiken, Roma Kaul, Takuro Fujiwara, Isaac S Harris, Tian Zhang, Steven P Gygi, Joan S Brugge
Cancer-associated mutations that stabilize NRF2, an oxidant defense transcription factor, are predicted to promote tumor development. Here, utilizing 3D cancer spheroid models coupled with CRISPR-Cas9 screens, we investigate the molecular pathogenesis mediated by NRF2 hyperactivation. NRF2 hyperactivation was necessary for proliferation and survival in lung tumor spheroids. Antioxidant treatment rescued survival but not proliferation, suggesting the presence of distinct mechanisms. CRISPR screens revealed that spheroids are differentially dependent on the mammalian target of rapamycin (mTOR) for proliferation and the lipid peroxidase GPX4 for protection from ferroptosis of inner, matrix-deprived cells...
December 3, 2020: Molecular Cell
#37
JOURNAL ARTICLE
Anna A Lauer, Janine Mett, Daniel Janitschke, Andrea Thiel, Christoph P Stahlmann, Cornel M Bachmann, Felix Ritzmann, Bianca Schrul, Ulrike C Müller, Reuven Stein, Matthias Riemenschneider, Heike S Grimm, Tobias Hartmann, Marcus O W Grimm
One of the major pathological hallmarks of Alzheimer´s disease (AD) is an accumulation of amyloid-β (Aβ) in brain tissue leading to formation of toxic oligomers and senile plaques. Under physiological conditions, a tightly balanced equilibrium between Aβ-production and -degradation is necessary to prevent pathological Aβ-accumulation. Here, we investigate the molecular mechanism how insulin-degrading enzyme (IDE), one of the major Aβ-degrading enzymes, is regulated and how amyloid precursor protein (APP) processing and Aβ-degradation is linked in a regulatory cycle to achieve this balance...
November 2020: Aging Cell
#38
JOURNAL ARTICLE
Qian Yang, Mei Wu, Ya-Li Zhu, Ya-Qiong Yang, Yan-Zhen Mei, Chuan-Chao Dai
Flavonoids, which are mainly extracted from plants, are important antioxidants and play an important role in human diseases. However, the growing market demand is limited by low productivity and complex production processes. Herein, the flavonoids biosynthesis pathway of the endophytic fungus Phomopsis liquidambaris was revealed. The mitogen-activated protein kinase kinase (MAPKK) of the strain was disrupted using a newly constructed CRISPR-Cas9 system mediated by two gRNAs which was conducive to cause plasmid loss...
January 2021: Biotechnology Letters
#39
JOURNAL ARTICLE
Yuchen Liu, Weiren Huang, Zhiming Cai
The logical AND gate gene circuit based on the CRISPR-Cas9 system can distinguish bladder cancer cells from normal bladder epithelial cells. However, the layered artificial gene circuits have the problems of high complexity, difficulty in accurately predicting the behavior, and excessive redundancy, which cannot be applied to clinical translation. Here, we construct minigene circuits based on the CRISPReader, a technology used to control promoter-less gene expression in a robust manner. The minigene circuits significantly induce robust gene expression output in bladder cancer cells, but have nearly undetectable gene expression in normal bladder epithelial cells...
October 30, 2020: Nature Communications
#40
JOURNAL ARTICLE
Jinliang Gao, Lin Wu, Daidi Yang, Weida Gong, Jinke Wang
The CRISPR/Cas9 has high specificity to its target DNA as a gene editing tool. This characteristic makes it useful for DNA detection. Combining the advantages of CRISPR/Cas9 and PCR, this study established a novel CRISPR/Cas9-based DNA detection method, named as CRISPR/Cas9-typing PCR version 4.0 (ctPCR4.0). This method can detect target DNA in one spot with high specificity and sensitivity. In a homogenous reaction, the target DNA is first cleaved by a pair of Cas9-sgRNA complexes and thus releases two single strands with free 3' ends, allowing a pair of oligonucleotides to anneal with...
October 27, 2020: Journal of Molecular Diagnostics: JMD
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