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https://www.readbyqxmd.com/read/28643252/generation-of-knock-in-mouse-by-genome-editing
#1
Wataru Fujii
Knock-in mice are useful for evaluating endogenous gene expressions and functions in vivo. Instead of the conventional gene-targeting method using embryonic stem cells, an exogenous DNA sequence can be inserted into the target locus in the zygote using genome editing technology. In this chapter, I describe the generation of epitope-tagged mice using engineered endonuclease and single-stranded oligodeoxynucleotide through the mouse zygote as an example of how to generate a knock-in mouse by genome editing.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28643250/genome-editing-in-mouse-zygotes-and-embryonic-stem-cells-by-introducing-sgrna-cas9-expressing-plasmids
#2
Taichi Noda, Asami Oji, Masahito Ikawa
In mammalian cells, genome editing with the single guide RNA (sgRNA)/Cas9 complex allows for high targeting efficiency within a relatively short time frame with the added benefits of being low cost and easy to design. sgRNA/Cas9-mediated editing in mouse zygotes has accelerated the analysis of gene functions and the generation of mouse models of human diseases. Despite the benefits, this method still suffers from several problems, such as mosaicism in the founder generation which complicates genotyping and phenotypical analyses, and the low efficiency of more complicated genome editing...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28643089/time-lapse-observation-and-transcriptome-analysis-of-a-case-with-repeated-multiple-pronuclei-after-ivf-icsi
#3
J Dai, L Z Leng, C F Lu, F Gong, S P Zhang, W Zheng, G X Lu, G Lin
PURPOSE: The purpose of this study was to investigate the cause of repeated multipronucleus (MPN) formation in zygotes in a patient after both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). METHOD: This is a case study. A patient had unexplained primary infertility with recurring total MPN zygotes after IVF and ICSI cycles. Time-lapse monitoring of pronucleus formation was carried out. Embryos developed from MPN zygotes were analyzed by fluorescence in situ hybridization (FISH)...
June 22, 2017: Journal of Assisted Reproduction and Genetics
https://www.readbyqxmd.com/read/28639312/quantification-of-transmission-risk-in-a-male-patient-with-a-flnb-mosaic-mutation-causing-larsen-syndrome-implications-for-genetic-counselling-in-post-zygotic-mosaicism-cases
#4
Marie Bernkopf, David Hunt, Nils Koelling, Tim Morgan, Amanda L Collins, Joanna Fairhurst, Stephen P Robertson, Andrew G L Douglas, Anne Goriely
We report the case of a male patient with Larsen syndrome found to be mosaic for a novel point mutation in FLNB in whom it was possible to provide evidence-based personalised counselling on transmission risk to future offspring. Using dideoxy-sequencing, a low-level FLNB c.698A>G, encoding p.(Tyr233Cys) mutation was detected in buccal mucosa and fibroblast DNA. Mutation quantification was performed by deep Next-Generation sequencing (NGS) of DNA extracted from three somatic tissues (blood, fibroblasts, saliva) and a sperm sample...
June 21, 2017: Human Mutation
https://www.readbyqxmd.com/read/28637346/an-ion-beam-induced-arabidopsis-mutant-with-marked-chromosomal-rearrangement
#5
Ayako N Sakamoto, Vo Thi Thuong Lan, Satoru Fujimoto, Sachihiro Matsunaga, Atsushi Tanaka
Ion beams have been used as an effective tool in mutation breeding for the creation of crops with novel characteristics. Recent analyses have revealed that ion beams induce large chromosomal alterations, in addition to small mutations comprising base changes or frameshifts. In an effort to understand the potential capability of ion beams, we analyzed an Arabidopsis mutant possessing an abnormal genetic trait. The Arabidopsis mutant uvh3-2 is hypersensitive to UVB radiation when photoreactivation is unavailable...
June 15, 2017: Journal of Radiation Research
https://www.readbyqxmd.com/read/28637085/successful-delivery-derived-from-cryopreserved-rare-human-spermatozoa-with-novel-cryopiece
#6
J Sun, W Chen, L Zhou, J Hu, Z Li, Z Zhang, Y Wu
Herein, we report the clinical outcomes following intracytoplasmic sperm injection (ICSI) with cryopiece cryopreserved rare human spermatozoa from severe male factor infertility patients. We established a novel cryopiece system on the basis of previous studies. In this study, 126 spermatozoa from four patients with non-obstructiveazoospermia (NOA) or severe oligozoospermia were stored in cryopiece and then thawed on the day of the oocyte retrieval, 88 (83%) spermatozoa were recovered with a 47.5% (38 of 80) motile rate...
June 21, 2017: Andrology
https://www.readbyqxmd.com/read/28634273/the-c-elegans-vapb-homolog-vpr-1-is-a-permissive-signal-for-gonad-development
#7
Pauline A Cottee, Tim Cole, Jessica Schultz, Hieu D Hoang, Jack Vibbert, Sung Min Han, Michael A Miller
VAMP/synaptobrevin-associated proteins (VAPs) contain an N-terminal major sperm protein domain (MSPd) that is associated with amyotrophic lateral sclerosis. VAPs have an intracellular housekeeping function, as well as an extracellular signaling function mediated by the secreted MSPd. Here we show that the C. elegans VAP homolog VPR-1 is essential for gonad development. vpr-1 null mutants are maternal effect sterile due to arrested gonadogenesis following embryo hatching. Somatic gonadal precursor cells and germ cells fail to proliferate fully and complete their respective differentiation programs...
June 15, 2017: Development
https://www.readbyqxmd.com/read/28631407/zygote-arrest-3-that-encodes-the-trna-ligase-is-essential-for-zygote-division-in-arabidopsis
#8
Ke-Jin Yang, Lei Guo, Xiu-Li Hou, Hua-Qin Gong, Chun-Ming Liu
In sexual organisms, division of the zygote initiates a new life cycle. Although several genes involved in zygote division are known in plants, how the zygote is activated to start embryogenesis remains elusive. Here, we showed that a mutation in ZYGOTE-ARREST 3 (ZYG3) in Arabidopsis led to a tight zygote-lethal phenotype. Map-based cloning revealed that ZYG3 encodes the tRNA ligase AtRNL, which is a single-copy gene in the Arabidopsis genome. Expression analyses showed that AtRNL is expressed throughout zygotic embryogenesis, and in meristematic tissues...
June 20, 2017: Journal of Integrative Plant Biology
https://www.readbyqxmd.com/read/28630100/nlrp2-and-faf1-deficiency-blocks-early-embryogenesis-in-the-mouse
#9
Hui Peng, Hai-Jun Liu, Fang Liu, Yuyun Gao, Jing Chen, Jianchao Huo, Jinglin Han, Tianfang Xiao, Wenchang Zhang
Nlrp2 is a maternal effect gene specifically expressed by mouse ovaries; deletion of this gene from zygotes is known to result in early embryonic arrest. In the present study, we identified FAF1 protein as a specific binding partner of the NLRP2 protein in both mouse oocytes and preimplantation embryos. In addition to early embryos, both Faf1 mRNA and protein were detected in multiple tissues. NLRP2 and FAF1 proteins were co-localized to both the cytoplasm and nucleus during the development of oocytes and preimplantation embryos...
June 19, 2017: Reproduction: the Official Journal of the Society for the Study of Fertility
https://www.readbyqxmd.com/read/28629791/inositol-phosphate-signaling-as-mediator-for-growth-and-sexual-reproduction-in-podospora-anserina
#10
Ning Xie, Gwenaël Ruprich-Robert, Florence Chapeland-Leclerc, Evelyne Coppin, Hervé Lalucque, Sylvain Brun, Robert Debuchy, Philippe Silar
The molecular pathways involved in the development of multicellular fruiting bodies in fungi are still not well known. Especially, the interplay between the mycelium, the female tissues and the zygotic tissues of the fruiting bodies is poorly documented. Here, we describe PM154, a new strain of the model ascomycetes Podospora anserina able to mate with itself and that enabled the easy recovery of new mutants affected in fruiting body development. By complete genome sequencing of spod1, one of the new mutants, we identified an inositol phosphate polykinase gene as essential, especially for fruiting body development...
June 16, 2017: Developmental Biology
https://www.readbyqxmd.com/read/28629378/microrna-130b-is-involved-in-bovine-granulosa-and-cumulus-cells-function-oocyte-maturation-and-blastocyst-formation
#11
Pritam Bala Sinha, Dawit Tesfaye, Franca Rings, Munir Hossien, Michael Hoelker, Eva Held, Christaine Neuhoff, Ernst Tholen, Karl Schellander, Dessie Salilew-Wondim
BACKGROUND: Oocyte maturation and preimplantation embryo development are controlled by array of genes that are post-transcriptionally regulated by microRNAs. With respect to this, previously, we identified altered expression of microRNA-130b (miR-130b) during oocyte maturation. Here, we aimed to investigate the role of miR-130b in bovine granulosa and cumulus cell function, oocyte maturation and preimplantation embryo development using gain- and loss-of- function approach. METHODS: For this study, the granulosa cells, cumulus cells and the oocytes were collected from ovaries obtained from slaughterhouse...
June 19, 2017: Journal of Ovarian Research
https://www.readbyqxmd.com/read/28626639/a-non-mosaic-porcn-mutation-in-a-male-with-severe-congenital-anomalies-overlapping-focal-dermal-hypoplasia
#12
Simran Madan, Wei Liu, James T Lu, V Reid Sutton, Bryant Toth, Priscilla Joe, John R Waterson, Richard A Gibbs, Ignatia B Van den Veyver, Edward J Lammer, Philippe M Campeau, Brendan H Lee
Mutations in the PORCN gene cause the X-linked dominant condition focal dermal hypoplasia (FDH). Features of FDH include striated pigmentation of the skin, ocular and skeletal malformations. FDH is generally associated with in utero lethality in non-mosaic males and most of the currently reported male patients show mosaicism due to de novo post-zygotic mutations in the PORCN gene. There is only one previous report of a surviving male with an inherited mutation in the PORCN gene. Here, we report two male siblings with multiple malformations including skeletal, ocular and renal defects overlapping with FDH...
September 2017: Molecular Genetics and Metabolism Reports
https://www.readbyqxmd.com/read/28624889/transcriptional-control-of-unequal-cleavage-in-early-tubifex-embryos
#13
Momoe Aoki, Takashi Shimizu
Early embryos of the clitellate annelid Tubifex (oligochaete) undergo a series of unequal spiral cell divisions before the descendants of the D quadrant micromeres (cells 2d and 4d) divide bilaterally. Here, we show that inhibition of zygotic transcription by microinjection of α-amanitin (transcription inhibitor) exclusively converts unequal cleavage in cell 2d(11) (granddaughter of 2d) into equal cleavage while other unequal cleavages and ensuing bilateral cleavages in cells 4d and 2d(111) (great-granddaughter of 2d) all proceed in a normal fashion in the presence of this inhibitor...
June 17, 2017: Development Genes and Evolution
https://www.readbyqxmd.com/read/28622964/grand-paternal-inheritance-of-x-linked-myotubular-myopathy-due-to-mosaicism-and-identification-of-necklace-fibers-in-an-asymptomatic-male
#14
Carola Hedberg-Oldfors, Kittichate Visuttijai, Alexandra Topa, Mar Tulinius, Anders Oldfors
X-linked recessive myotubular myopathy (XLMTM) is a disorder associated with mutations in the myotubularin gene (MTM1) that usually affects boys, with transmission of the mutated allele from the mother. Here we describe a family with unexpected grand paternal transmission of a novel mutation in MTM1 (c.646_648dupGTT; p.Val216dup) identified in a severely affected infant boy with a centronuclear myopathy. We confirmed the carrier status of the mother, but surprisingly we found that her father was a carrier of the mutated MTM1 gene together with wild-type MTM1...
May 10, 2017: Neuromuscular Disorders: NMD
https://www.readbyqxmd.com/read/28621432/regulatory-heterochronies-and-loose-temporal-scaling-between-sea-star-and-sea-urchin-regulatory-circuits
#15
Tsvia Gildor, Veronica Hinman, Smadar Ben-Tabou-De-Leon
It has long been argued that heterochrony, a change in relative timing of a developmental process, is a major source of evolutionary innovation. Heterochronic changes of regulatory gene activation could be the underlying molecular mechanism driving heterochronic changes through evolution. Here, we compare the temporal expression profiles of key regulatory circuits between sea urchin and sea star, representative of two classes of Echinoderms that shared a common ancestor about 500 million years ago. The morphologies of the sea urchin and sea star embryos are largely comparable, yet, differences in certain mesodermal cell types and ectodermal patterning result in distinct larval body plans...
2017: International Journal of Developmental Biology
https://www.readbyqxmd.com/read/28621329/single-cell-multi-omics-sequencing-of-mouse-early-embryos-and-embryonic-stem-cells
#16
Fan Guo, Lin Li, Jingyun Li, Xinglong Wu, Boqiang Hu, Ping Zhu, Lu Wen, Fuchou Tang
Single-cell epigenome sequencing techniques have recently been developed. However, the combination of different layers of epigenome sequencing in an individual cell has not yet been achieved. Here, we developed a single-cell multi-omics sequencing technology (single-cell COOL-seq) that can analyze the chromatin state/nucleosome positioning, DNA methylation, copy number variation and ploidy simultaneously from the same individual mammalian cell. We used this method to analyze the reprogramming of the chromatin state and DNA methylation in mouse preimplantation embryos...
June 16, 2017: Cell Research
https://www.readbyqxmd.com/read/28619824/the-histone-3-lysine-4-methyltransferase-setd1b-is-a-maternal-effect-gene-required-for-the-oogenic-gene-expression-program
#17
David Brici, Qinyu Zhang, Susanne Reinhardt, Andreas Dahl, Hella Hartmann, Kerstin Schmidt, Neha Goveas, Jiahao Huang, Lenka Gahurova, Gavin Kelsey, Konstantinos Anastassiadis, A Francis Stewart, Andrea Kranz
Germ cell development involves major reprogramming of the epigenome to prime the zygote for totipotency. Histone 3 lysine 4 (H3K4) methylations are universal epigenetic marks mediated in mammals by six H3K4 methyltransferases related to fly Trithorax, including two yeast Set1 orthologs: Setd1a and Setd1b. Whereas Setd1a plays no role in oogenesis, we report that Setd1b deficiency causes female sterility. Oocyte specific Gdf9iCre conditional knockout (Setd1b(Gdf9) cKO) ovaries develop through all stages however follicular loss accumulated with age and unfertilized metaphase II (MII) oocytes exhibited irregularities of the zona pellucida and meiotic spindle...
June 15, 2017: Development
https://www.readbyqxmd.com/read/28619362/peroxiredoxin-1-from-cuttlefish-sepiella-maindroni-molecular-characterization-of-development-and-its-immune-response-against-vibrio-alginolyticus
#18
Weiwei Song, Changkao Mu, Ronghua Li, Chunlin Wang
The aim of this work was constructive to understand the function of peroxiredoxin (PRDX) family member Peroxiredoxin 1 in Sepiella maindroni (SmPrx1) through molecular mechanisms of reproduction, embryonic development and immune responses to Vibrio alginolyticus. The full-length cDNA of SmPrx1 was of 1062 bp, contains a 5' untranslated region (UTR) of 79bp, a 3' UTR of 359 bp, an open reading frame of 624 bp encoding 207 amino acids. The conserved peroxidase catalytic center "FYPLDFTFVCPTEI" and "GEVCPA" were observed in the sequence of SmPrx1; this indicated that it was a member of 2-Cys Prx...
June 12, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28615321/par-proteins-regulate-maintenance-phase-myosin-dynamics-during-c-elegans-zygote-polarization
#19
Lawrence E Small, Adriana T Dawes
Establishment of anterior-posterior polarity in the C. elegans zygote requires two different processes: mechanical activity of the actin-myosin cortex and biochemical activity of partitioning defective (PAR) proteins. Here, we have analyzed how PARs regulate the behavior of the cortical motor protein non-muscle myosin (NMY-2) to complement recent efforts that investigate how PARs regulate the Rho GTPase CDC-42, which in turn regulates the actin-myosin cortex. We found that PAR-3 and PAR-6 concentrate CDC-42-dependent NMY-2 in the anterior cortex, while PAR-2 inhibits CDC-42-dependent NMY-2 in the posterior by inhibiting PAR-3 and PAR-6...
June 14, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28615282/maternal-haploid-a-metalloprotease-enriched-at-the-largest-satellite-repeat-and-essential-for-genome-integrity-in-drosophila-embryos
#20
Xiaona Tang, Jingguo Cao, Liang Zhang, Yingzi Huang, Qianyi Zhang, Yikang S Rong
The incorporation of the paternal genome into the zygote during fertilization requires chromatin remodeling. The maternal haploid (mh) mutation in Drosophila affects this process and leads to the formation of haploid embryos without the paternal genome. mh encodes the Drosophila homolog of SPRTN, a conserved protease essential for resolving DNA-protein cross-linked products. Here we characterize the role of MH in genome maintenance. It is not understood how MH protects the paternal genome during fertilization particularly in lights of our finding that MH is present in both parental pro-nuclei during zygote formation...
June 14, 2017: Genetics
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