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https://www.readbyqxmd.com/read/28273380/an-aanat-asmt-transgenic-animal-model-constructed-with-crispr-cas9-system-serving-as-the-mammary-gland-bioreactor-to-produce-melatonin-enrich-milk-in-sheep
#1
Teng Ma, Jingli Tao, Minghui Yang, Changjiu He, Xiuzhi Tian, Xiaosheng Zhang, Jinlong Zhang, Shoulong Deng, Jianzhong Feng, Zhenzhen Zhang, Jing Wang, Pengyun Ji, Yukun Song, Pingli He, Hongbing Han, Juncai Fu, Zhengxing Lian, Guoshi Liu
Melatonin as a potent antioxidant exhibits important nutritional and medicinal values. To produce melatonin-enriching milk will benefit to the consumers. In this study, a sheep bioreactor which generates melatonin-enriching milk has been successfully developed by the technology that combined CRISPR/Cas9 system and microinjection. The AANAT and ASMT were cloned from pineal gland of Dorper sheep (Ovis aries). The in vitro studies found that AANAT and ASMT were successfully transferred to the mammary epithelial cell lines and significantly increased melatonin production in the culture medium compared to the non-transgenic cell lines...
March 8, 2017: Journal of Pineal Research
https://www.readbyqxmd.com/read/28268229/the-hdac-inhibitor-laq824-enhances-epigenetic-reprogramming-and-in-vitro-development-of-porcine-scnt-embryos
#2
Jun-Xue Jin, Sanghoon Lee, Anukul Taweechaipaisankul, Geon A Kim, Byeong Chun Lee
BACKGROUND/AIMS: Hypoacetylation caused by aberrant epigenetic nuclear reprogramming results in low efficiency of mammalian somatic cell nuclear transfer (SCNT). Many epigenetic remodeling drugs have been used in attempts to improve in vitro development of porcine SCNT embryos. In this study, we examined the effects of LAQ824, a structurally novel histone acetylase inhibitor, on the nuclear reprogramming and in vitro development of porcine SCNT embryos. METHODS: LAQ824 treatment was supplemented during the culture of SCNT embryos...
March 7, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28251317/crispr-cas9-mediated-gene-editing-in-human-zygotes-using-cas9-protein
#3
Lichun Tang, Yanting Zeng, Hongzi Du, Mengmeng Gong, Jin Peng, Buxi Zhang, Ming Lei, Fang Zhao, Weihua Wang, Xiaowei Li, Jianqiao Liu
Previous works using human tripronuclear zygotes suggested that the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system could be a tool in correcting disease-causing mutations. However, whether this system was applicable in normal human (dual pronuclear, 2PN) zygotes was unclear. Here we demonstrate that CRISPR/Cas9 is also effective as a gene-editing tool in human 2PN zygotes. By injection of Cas9 protein complexed with the appropriate sgRNAs and homology donors into one-cell human embryos, we demonstrated efficient homologous recombination-mediated correction of point mutations in HBB and G6PD...
March 1, 2017: Molecular Genetics and Genomics: MGG
https://www.readbyqxmd.com/read/28224725/quisinostat-treatment-improves-histone-acetylation-and-developmental-competence-of-porcine-somatic-cell-nuclear-transfer-embryos
#4
Long Jin, Qing Guo, Hai-Ying Zhu, Xiao-Xu Xing, Guang-Lei Zhang, Mei-Fu Xuan, Qi-Rong Luo, Zhao-Bo Luo, Jun-Xia Wang, Xi-Jun Yin, Jin-Dan Kang
Abnormal epigenetic modifications are considered a main contributing factor to low cloning efficiency. In the present study, we explored the effects of quisinostat, a novel histone deacetylase inhibitor, on blastocyst formation rate in porcine somatic-cell nuclear transfer (SCNT) embryos, on acetylation of histone H3 lysine 9 (AcH3K9), and on expression of POU5F1 protein and apoptosis-related genes BAX and BCL2. Our results showed that treatment with 10 nM quisinostat for 24 h significantly improved the development of reconstructed embryos compared to the untreated group (19...
February 22, 2017: Molecular Reproduction and Development
https://www.readbyqxmd.com/read/28222427/impact-of-fmr1-pre-mutation-status-on-blastocyst-development-in-patients-undergoing-pre-implantation-genetic-diagnosis
#5
Anne P Hutchinson, Nigel Pereira, Debra P Lilienthal, Siobhán Coveney, Jovana P Lekovich, Rony T Elias, Zev Rosenwaks
BACKGROUND/AIMS: The study aimed to investigate the impact of fragile X mental retardation 1 (FMR1) pre-mutation status on blastocyst development in patients undergoing pre-implantation genetic diagnosis (PGD). METHODS: Case-control study of patients <40 years undergoing PGD at blastocyst stage for FMR1 pre-mutation status. Age-matched patients undergoing PGD for other single gene disorders were considered controls. Blastocyst development, calculated per metaphase II (MII) oocyte retrieved and per 2 pronuclear (2PN) embryos, was compared between the 2 groups...
February 22, 2017: Gynecologic and Obstetric Investigation
https://www.readbyqxmd.com/read/28152037/reassessing-the-feasibility-of-the-zygote-score-for-predicting-embryo-viability-in-ivf-icsi-using-the-gnrh-antagonist-protocol-compared-to-the-long-protocol
#6
Pin-Yao Lin, Fu-Jen Huang, Fu-Tsai Kung, Yi-Chi Lin, Hsin-Ju Chiang, Yu-Ju Lin, Kuo-Chung Lan
BACKGROUND: Many factors from the oocyte/sperm or the process of fertilization may affect the zygote formation. The zygote score (Z-score) describes the quality of a human zygote based on its pronuclear morphology, nucleolar precursor bodies, and alignment of polar bodies, and it can be used in the selection process at the zygote stage for embryo transfer or cryopreservation. OBJECTIVE: The aim of this retrospective cohort study was to investigate the relationship between different controlled ovarian stimulation (COS) protocols and the zygote score (Z-score) and to assess the feasibility of the Z-score for predicting embryo survival in the GnRH-antagonist (GnRH-ant) protocol...
2017: PloS One
https://www.readbyqxmd.com/read/28111439/ascorbic-acid-increases-demethylation-in-somatic-cell-nuclear-transfer-embryos-of-the-pig-sus-scrofa
#7
Minghui Zhao, Tai-Young Hur, Jingu No, Yoonseok Nam, Hyeunkyu Kim, Gi-Sun Im, Seunghoon Lee
Objective: Investigated the effect and mechanism of ascorbic acid on the development of porcine embryos produced by somatic cell nuclear transfer (SCNT). Methods: Porcine embryos were produced by SCNT and cultured in the presence or absence of ascorbic acid. TET3 in oocytes was knocked down by siRNA injection. After ascorbic acid treatment, reprogramming genes were analyzed by realtime RT-PCR. Furthermore, relative 5mC and 5hmC content in pronuclear were detected by realtime PCR...
January 13, 2017: Asian-Australasian Journal of Animal Sciences
https://www.readbyqxmd.com/read/28087350/corrigendum-to-pregnancy-derived-from-human-zygote-pronuclear-transfer-in-a-patient-who-had-arrested-embryos-after-ivf-reproductive-biomedicine-online-33-2016-529-533
#8
John Zhang, Guanglun Zhuang, Yong Zeng, Jamie Grifo, Carlo Acosta, Yimin Shu, Hui Liu
No abstract text is available yet for this article.
January 10, 2017: Reproductive Biomedicine Online
https://www.readbyqxmd.com/read/28053091/fumarylacetoacetate-hydrolase-knockout-rabbit-model-for-hereditary-tyrosinemia-type-1
#9
Li Li, Quanjun Zhang, Huaqiang Yang, Qingjian Zou, Chengdan Lai, Fei Jiang, Ping Zhao, Zhiwei Luo, Jiayin Yang, Qian Chen, Yan Wang, Philip N Newsome, Jon Frampton, Patrick H Maxwell, Wenjuan Li, Shuhan Chen, Dongye Wang, Tak-Shing Siu, Sidney Tam, Hung-Fat Tse, Baoming Qin, Xichen Bao, Miguel A Esteban, Liangxue Lai
Hereditary tyrosinemia type 1 (HT1) is a severe human autosomal recessive disorder caused by the deficiency of fumarylacetoacetate hydroxylase (FAH), an enzyme catalyzing the last step in the tyrosine degradation pathway. Lack of FAH causes accumulation of toxic metabolites (fumarylacetoacetate and succinylacetone) in blood and tissues, ultimately resulting in severe liver and kidney damage with onset that ranges from infancy to adolescence. This tissue damage is lethal but can be controlled by administration of 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedion (NTBC), which inhibits tyrosine catabolism upstream of the generation of fumarylacetoacetate and succinylacetone...
January 4, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28050628/epigenetic-modifications-and-reprogramming-in-paternal-pronucleus-sperm-preimplantation-embryo-and-beyond
#10
REVIEW
Yuki Okada, Kosuke Yamaguchi
Pronuclear/zygotic stage is the very first stage of life. In this period, paternal pronucleus undergoes massive chromatin remodeling called "paternal reprogramming" including protamine-histone replacement and subsequent acquisition of epigenetic modifications. Although these consecutive events are required for the initiation of maternal-zygotic transition, the precise role of paternal reprogramming and its effect on subsequent embryonic development has been largely unknown to date. Recently, various new techniques, especially next-generation sequencing (NGS) and RNAi microinjection contribute to unveil the epigenetic transition from both paternal and maternal to early preimplantation embryos, suggesting not only the simple transcriptional regulation by transcription factors but also dynamic structural alteration of chromatin to initiate the wave of zygotic gene transcription...
January 3, 2017: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/28031247/thioredoxin-dependent-disulfide-bond-reduction-is-required-for-protamine-eviction-from-sperm-chromatin
#11
Alexander V Emelyanov, Dmitry V Fyodorov
Cysteine oxidation in protamines leads to their oligomerization and contributes to sperm chromatin compaction. Here we identify the Drosophila thioredoxin Deadhead (DHD) as the factor responsible for the reduction of intermolecular disulfide bonds in protamines and their eviction from sperm during fertilization. Protamine chaperone TAP/p32 dissociates DNA-protamine complexes in vitro only when protamine oligomers are first converted to monomers by DHD. dhd-null embryos cannot decondense sperm chromatin and terminate development after the first pronuclear division...
December 15, 2016: Genes & Development
https://www.readbyqxmd.com/read/28018848/mitochondrial-transfer-implications-for-assisted-reproductive-technologies
#12
REVIEW
A S Reznichenko, C Huyser, M S Pepper
The use of mitochondrial transfer as a clinic procedure is drawing closer to reality. Here we provide a detailed overview of mitochondrial transfer techniques - both established and recent - including pronuclear, spindle, ooplasmic and blastomere transfer. Reasons as to why some techniques are more suitable for the prevention of mitochondrial DNA disease than others, as well as the advantages and disadvantages of each methodology, are discussed. The possible clinical introduction of these techniques has raised concerns about the adverse effects they may have on resultant embryos and offspring...
December 2016: Applied & Translational Genomics
https://www.readbyqxmd.com/read/27933305/robust-generation-of-transgenic-mice-by-simple-hypotonic-solution-mediated-delivery-of-transgene-in-testicular-germ-cells
#13
Abul Usmani, Nirmalya Ganguli, Subodh K Jain, Nilanjana Ganguli, Rajesh Kumar Sarkar, Mayank Choubey, Mansi Shukla, Hironmoy Sarkar, Subeer S Majumdar
Our ability to decipher gene sequences has increased enormously with the advent of modern sequencing tools, but the ability to divulge functions of new genes have not increased correspondingly. This has caused a remarkable delay in functional interpretation of several newly found genes in tissue and age specific manner, limiting the pace of biological research. This is mainly due to lack of advancements in methodological tools for transgenesis. Predominantly practiced method of transgenesis by pronuclear DNA-microinjection is time consuming, tedious, and requires highly skilled persons for embryo-manipulation...
2016: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/27878881/beneficial-effect-of-extracellular-adenosine-5-triphosphate-treatment-on-the-indochinese-leopard-panthera-pardus-delacouri-sperm-quality-after-cryopreservation
#14
P Thuwanut, W Tipkantha, B Siriaroonrat, P Comizzoli, K Chatdarong
The Indochinese leopard (Panthera pardus delacouri) population, included in CITES Appendix I, has been declining for decades. Proper gamete preservation condition is critical for breeding programme management using artificial insemination or in vitro fertilization (IVF). The present study aimed at investigating the impact of post-thawing treatment of leopard semen with extracellular adenosine 5'-triphosphate (ATPe) on sperm quality (including morphological traits and ability to fertilize an oocyte). Semen from six adult male leopards was collected by electroejaculation (one ejaculation per cat)...
November 22, 2016: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/27833197/ethical-aspects-of-nuclear-and-mitochondrial-dna-transfer
#15
José Rafael Blesa, Julio Tudela, Justo Aznar
Somatic cell nuclear transfer (SCNT) (cloning), as a reproductive or therapeutic method, and mitochondrial DNA transfer, as a method to prevent the transmission of mitochondrial diseases, are analyzed in this paper from a bioethics perspective. The licit purpose of being able to treat certain diseases, as in the case of SCNT, cannot justify, in any case, resorting to illicit means such as the manipulation, selection, and elimination of human embryos in the blastocyst phase, by using cell lines obtained from them...
May 2016: Linacre Quarterly
https://www.readbyqxmd.com/read/27827818/oocyte-embryo-and-blastocyst-cryopreservation-in-art-systematic-review-and-meta-analysis-comparing-slow-freezing-versus-vitrification-to-produce-evidence-for-the-development-of-global-guidance
#16
REVIEW
Laura Rienzi, Clarisa Gracia, Roberta Maggiulli, Andrew R LaBarbera, Daniel J Kaser, Filippo M Ubaldi, Sheryl Vanderpoel, Catherine Racowsky
BACKGROUND: Successful cryopreservation of oocytes and embryos is essential not only to maximize the safety and efficacy of ovarian stimulation cycles in an IVF treatment, but also to enable fertility preservation. Two cryopreservation methods are routinely used: slow-freezing or vitrification. Slow-freezing allows for freezing to occur at a sufficiently slow rate to permit adequate cellular dehydration while minimizing intracellular ice formation. Vitrification allows the solidification of the cell(s) and of the extracellular milieu into a glass-like state without the formation of ice...
November 8, 2016: Human Reproduction Update
https://www.readbyqxmd.com/read/27788132/maintenance-of-xist-imprinting-depends-on-chromatin-condensation-state-and-rnf12-dosage-in-mice
#17
Atsushi Fukuda, Atsushi Mitani, Toshiyuki Miyashita, Takashi Sado, Akihiro Umezawa, Hidenori Akutsu
In female mammals, activation of Xist (X-inactive specific transcript) is essential for establishment of X chromosome inactivation. During early embryonic development in mice, paternal Xist is preferentially expressed whereas maternal Xist (Xm-Xist) is silenced. Unlike autosomal imprinted genes, Xist imprinting for Xm-Xist silencing was erased in cloned or parthenogenetic but not fertilized embryos. However, the molecular mechanism underlying the variable nature of Xm-Xist imprinting is poorly understood. Here, we revealed that Xm-Xist silencing depends on chromatin condensation states at the Xist/Tsix genomic region and on Rnf12 expression levels...
October 2016: PLoS Genetics
https://www.readbyqxmd.com/read/27759163/generation-of-a-transgenic-medaka-oryzias-latipes-strain-for-visualization-of-nuclear-dynamics-in-early-developmental-stages
#18
Takanobu Inoue, Atsuo Iida, Shingo Maegawa, Atsuko Sehara-Fujisawa, Masato Kinoshita
In this study, we verified nuclear transport activity of an artificial nuclear localization signal (aNLS) in medaka fish (Oryzias latipes). We generated a transgenic medaka strain expresses the aNLS tagged enhanced green fluorescent protein (EGFP) driven by a medaka beta-actin promoter. The aNLS-EGFP was accumulated in the nuclei of somatic tissues and yolk nuclei of oocytes, but undetectable in the spermatozoa. The fluorescent signal was observed from immediately after fertilization by a maternal contribution...
December 2016: Development, Growth & Differentiation
https://www.readbyqxmd.com/read/27742403/effect-of-histone-acetylation-modification-with-mgcd0103-a-histone-deacetylase-inhibitor-on-nuclear-reprogramming-and-the-developmental-competence-of-porcine-somatic-cell-nuclear-transfer-embryos
#19
Long Jin, Hai-Ying Zhu, Qing Guo, Xiao-Chen Li, Yu-Chen Zhang, Cheng-Du Cui, Wen-Xue Li, Zheng-Yun Cui, Xi-Jun Yin, Jin-Dan Kang
Cloning remains as an important technique to enhance the reconstitution and distribution of animal population with high-genetic merit. One of the major detrimental factors of this technique is the abnormal epigenetic modifications. MGCD0103 is known as a histone deacetylase inhibitor. In this study, we investigated the effect of MGCD0103 on the in vitro blastocyst formation rate in porcine somatic cell nuclear transferred (SCNT) embryos and expression in acetylation of the histone H3 lysine 9 and histone H4 lysine 12...
January 1, 2017: Theriogenology
https://www.readbyqxmd.com/read/27733624/stronger-net-posterior-cortical-forces-and-asymmetric-microtubule-arrays-produce-simultaneous-centration-and-rotation-of-the-pronuclear-complex-in-the-early-caenorhabditis-elegans-embryo
#20
Valerie C Coffman, Matthew B A McDermott, Blerta Shtylla, Adriana T Dawes
Positioning of microtubule-organizing centers (MTOCs) incorporates biochemical and mechanical cues for proper alignment of the mitotic spindle and cell division site. Current experimental and theoretical studies in the early Caenorhabditis elegans embryo assume remarkable changes in the origin and polarity of forces acting on the MTOCs. These changes must occur over a few minutes, between initial centration and rotation of the pronuclear complex and entry into mitosis, and the models do not replicate in vivo timing of centration and rotation...
November 7, 2016: Molecular Biology of the Cell
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