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https://www.readbyqxmd.com/read/28643254/gene-targeting-in-rabbits-single-step-generation-of-knock-out-rabbits-by-microinjection-of-crispr-cas9-plasmids
#1
Yoshihiro Kawano, Arata Honda
The development of genome editing technology has allowed gene disruptions to be achieved in various animal species and has been beneficial to many mammals. Gene disruption using pluripotent stem cells is difficult to achieve in rabbits, but thanks to advances in genome editing technology, a number of gene disruptions have been conducted. This paper describes a simple and easy method for carrying out gene disruptions in rabbits using CRISPR/Cas9 in which the gene to be disrupted is marked, the presence or absence of off-target candidates is checked, and a plasmid allowing simultaneous expression of Cas9 and sgRNA is constructed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28643251/genome-editing-in-mouse-and-rat-by-electroporation
#2
Takehito Kaneko
Many knock-out/knock-in mouse and rat strains have been produced by genome editing techniques using engineered endonucleases, including zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), or clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9. Microinjection of engineered endonucleases into pronuclear-stage embryos is required to produce genome-edited rodents and the development of easy, rapid, and high-efficiency methods that do not require special skills such as microinjection is needed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28601410/total-number-of-oocytes-and-zygotes-are-predictive-of-live-birth-pregnancy-in-fresh-donor-oocyte-in%C3%A2-vitro-fertilization-cycles
#3
Eduardo Hariton, Keewan Kim, Sunni L Mumford, Marissa Palmor, Pietro Bortoletto, Eden R Cardozo, Anatte E Karmon, Mary E Sabatini, Aaron K Styer
OBJECTIVE: To evaluate the association of oocyte donor-recipient characteristics, oocyte donor response, and live birth pregnancy rate following fresh donor oocyte IVF-ET. DESIGN: Retrospective cohort study. SETTING: Academic reproductive medicine practice. PATIENT(S): Two hundred thirty-seven consecutive fresh donor oocyte IVF-ET cycles from January 1, 2007 to December 31, 2013 at the Massachusetts General Hospital Fertility Center...
June 7, 2017: Fertility and Sterility
https://www.readbyqxmd.com/read/28545999/a-comparative-analysis-of-the-efficacy-of-three-cryopreservation-protocols-on-the-survival-of-in-vitro-derived-cattle-embryos-at-pronuclear-and-blastocyst-stages
#4
Van Huong Do, Simon Walton, Sally Catt, Andrew W Taylor-Robinson
The effectiveness of three cryopreservation protocols (slow freezing, short equilibration vitrification and long equilibration vitrification) on in vitro-derived cattle embryos at expanded blastocyst and pronuclear stages was compared. 199 expanded blastocysts of good quality were assigned randomly into four treatment groups [control, non-cryopreserved (fresh, unfrozen); and the three cryopreservation methods]. The re-expansion of the cryopreserved blastocysts after 24 h in vitro culture was similar to that of the fresh control group...
May 22, 2017: Cryobiology
https://www.readbyqxmd.com/read/28475125/melatonin-promotes-the-in-vitro-development-of-microinjected-pronuclear-mouse-embryos-via-its-anti-oxidative-and-anti-apoptotic-effects
#5
Xiuzhi Tian, Feng Wang, Lu Zhang, Pengyun Ji, Jing Wang, Dongying Lv, Guangdong Li, Menglong Chai, Zhengxing Lian, Guoshi Liu
CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats) combined with pronuclear microinjection has become the most effective method for producing transgenic animals. However, the relatively low embryo developmental rate limits its application. In the current study, it was observed that 10(-7) M melatonin is considered an optimum concentration and significantly promoted the in vitro development of murine microinjected pronuclear embryos, as indicated by the increased blastocyst rate, hatching blastocyst rate and blastocyst cell number...
May 5, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28456971/use-of-histone-k-m-mutants-for-the-analysis-of-transcriptional-regulation-in-mouse-zygotes
#6
Keisuke Aoshima, Takashi Kimura, Yuki Okada
Histone modifications are dramatically altered during the pronuclear (PN) stage of zygotes, and more markedly in paternal than maternal pronuclei. Among various types of histone modifications, lysine methylation exhibits the most dynamic changes in the PN stage . To analyze the physiological functions of histone methylations, it is therefore important to elucidate the mechanism of epigenetic reprogramming. However, loss-of-function approaches using mutant histones whose lysine residues have been substituted with arginine residues are unable to erase histone modifications at all levels, since they are incapable of entirely replacing endogenous histones...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28448636/successful-vitrification-of-pronuclear-stage-pig-embryos-with-a-novel-cryoprotective-agent-carboxylated-%C3%AE%C2%B5-poly-l-lysine
#7
Maki Kamoshita, Tsubasa Kato, Katsuyoshi Fujiwara, Takafumi Namiki, Kazuaki Matsumura, Suong-Hyu Hyon, Junya Ito, Naomi Kashiwazaki
Vitrification is a powerful tool for the efficient production of offspring derived from cryopreserved oocytes or embryos in mammalian species including domestic animals. Genome editing technologies such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated (Cas)9 are now available even for domestic species, suggesting that the vitrification of embryos at the pronuclear stage (PN) will be more important because they could provide genomic host cells to be targeted by TALENs or CRISPR/Cas9...
2017: PloS One
https://www.readbyqxmd.com/read/28432708/in-vitro-production-of-porcine-zygotes-using-intracytoplasmic-injection-of-vitrified-sperm
#8
C C Arraztoa, C Baca Castex, G M Alvarez, P D Cetica, D M Neild
The objectives of this study were to evaluate if vitrified porcine spermatozoa are able to maintain their capacity to produce zygotes in vitro using intracytoplasmic sperm injection (ICSI) and to evaluate the zygote development in two in vitro atmospheric conditions: 5% CO2 and tri-gas. A group of porcine oocytes maturated in vitro were injected with vitrified-warmed sperm (treatment group) and another group, with sperm diluted and conserved at 17°C (control group). To evidence parthenogenetic activation, some oocytes were submitted to a Sham test...
April 21, 2017: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/28431828/clinical-use-of-monopronucleated-zygotes-following-blastocyst-culture-and-preimplantation-genetic-screening-including-verification-of-biparental-chromosome-inheritance
#9
Cara K Bradley, Maria Traversa, Natalie Hobson, Alison J Gee, Steven J McArthur
In assisted reproduction, embryos derived from monopronucleated (1PN) zygotes are considered abnormal and unsuitable for clinical use. Outcomes of 1PN-derived embryos designated for preimplantation genetic screening (PGS) were analysed. These embryos, especially from intracytoplasmic sperm injection (ICSI), were found to have a low developmental potential; 1PN and 2PN day 5 blastocyst development for IVF was 14.8% versus 36.4% (P < 0.0001), and for ICSI, 6.6% versus 34.0% (P < 0.0001), respectively. With the use of comparative genomic hybridization or next-generation sequencing, PGS was successfully carried out for 74 IVF and 32 ICSI 1PN-derived blastocysts, revealing adjusted abnormality rates of 39...
March 27, 2017: Reproductive Biomedicine Online
https://www.readbyqxmd.com/read/28429249/are-there-moral-differences-between-maternal-spindle-transfer-and-pronuclear-transfer
#10
César Palacios-González
This paper examines whether there are moral differences between the mitochondrial replacement techniques that have been recently developed in order to help women afflicted by mitochondrial DNA diseases to have genetically related children absent such conditions: maternal spindle transfer (MST) and pronuclear transfer (PNT). Firstly, it examines whether there is a moral difference between MST and PNT in terms of the divide between somatic interventions and germline interventions. Secondly, it considers whether PNT and MST are morally distinct under a therapy/creation optic...
April 20, 2017: Medicine, Health Care, and Philosophy
https://www.readbyqxmd.com/read/28419910/endometriosis-doubles-odds-for-miscarriage-in-patients-undergoing-ivf-or-icsi
#11
Corinna Pallacks, Jens Hirchenhain, Jan-Steffen Krüssel, Tanja N Fehm, Daniel Fehr
OBJECTIVE: To identify and estimate the importance of risk factors on pregnancy loss until the end of the second trimester after clinical pregnancy was achieved by either in vitro fertilisation (IVF) or intra-cytoplasmic sperm injection (ICSI). STUDY DESIGN: Retrospective cohort study including 588 cycles with fresh embryo transfers and 150 cycles with frozen-thawed embryo transfers using logistic regression. RESULTS: The rate of miscarriages subsequent to a fresh embryo transfer was significantly increased by a diagnosis of endometriosis (p=0...
June 2017: European Journal of Obstetrics, Gynecology, and Reproductive Biology
https://www.readbyqxmd.com/read/28412866/comparison-of-the-morphokinetic-parameters-of-embryos-according-to-ovarian-reserve-in-ivf-cycles
#12
Suleyman Akarsu, Funda Gode, Ahmet Zeki Isik, Hayriye Celenk, Ferda Burcu Tamer, Selcuk Erkilinc
The aim of this study is to evaluate the impact of ovarian reserve and age of women on early morphokinetic parameters of embryos with a time-lapse monitoring system. In total, 197 infertile couples with poor ovarian reserve (Group 1, n = 41), normal ovarian reserve (Group 2, n = 59), or polycystic ovaries (Group 3, n = 97) were included. The time from insemination to the following events were analyzed: pronuclear fading (Pnf) and cleavage to 2, 3, 4 and 5 cells. The optimal ranges for morphokinetic parameters of t5, s2 and cc2 in each group were also evaluated...
April 17, 2017: Gynecological Endocrinology
https://www.readbyqxmd.com/read/28380503/histone-deacetylase-inhibitor-m344-significantly-improves-nuclear-reprogramming-blastocyst-quality-and-in-vitro-developmental-capacity-of-cloned-pig-embryos
#13
L Jin, Q Guo, H-Y Zhu, X-X Xing, G-L Zhang, M-F Xuan, Q-R Luo, Z-B Luo, J-X Wang, H-M Choe, H-J Paek, X-J Yin, J-D Kang
M344 is a novel histone deacetylase inhibitor. There is no report on the effect of M344 treatment on the development of pig embryos after somatic cell nuclear transfer (SCNT). In the present study, we investigated the effect of M344 on the blastocyst formation rate in cloned embryos, acetylation level of histone H4 lysine 12 (AcH4K12), and the expression of pluripotency-related genes , , and . Our results indicated that treatment with 5 µ M344 for 6 h improved the development of porcine embryos, in comparison with the untreated group (25...
March 2017: Journal of Animal Science
https://www.readbyqxmd.com/read/28374747/mitochondrial-replacement-by-pre-pronuclear-transfer-in-human-embryos
#14
Keliang Wu, Tailai Chen, Sexin Huang, Cuiqing Zhong, Junhao Yan, Xiaoyu Zhang, Jinsong Li, Yuan Gao, Han Zhao, Zi-Jiang Chen
No abstract text is available yet for this article.
June 2017: Cell Research
https://www.readbyqxmd.com/read/28364522/spindlin-1-is-essential-for-metaphase-ii-stage-maintenance-and-chromosomal-stability-in-porcine-oocytes
#15
Jeong-Woo Choi, Ming-Hui Zhao, Shuang Liang, Jing Guo, Zi-Li Lin, Ying-Hua Li, Yu-Jin Jo, Nam-Hyung Kim, Xiang-Shun Cui
Study question: What is the function of Spindlin 1 (Spin1) in metaphase II stage oocytes in pigs? Summary answer: Depletion of Spin1 induces spontaneous oocyte activation and overexpression of Spin1 causes multinuclear formation through induction of DNA damage in porcine oocytes. What is known already: Little is known about the function of Spin1 in oocytes and embryos. In mouse oocytes, Spin1 is specifically expressed during gametogenesis and is essential for meiotic resumption...
March 1, 2017: Molecular Human Reproduction
https://www.readbyqxmd.com/read/28333185/randomized-open-trial-comparing-a-modified-double-lumen-needle-follicular-flushing-system-with-a-single-lumen-aspiration-needle-in-ivf-patients-with-poor-ovarian-response
#16
Kyra von Horn, Marion Depenbusch, Askan Schultze-Mosgau, Georg Griesinger
STUDY QUESTION: Is a modified double-lumen aspiration needle system with follicular flushing able to increase the mean oocyte yield by at least one in poor response IVF patients as compared to single-lumen needle aspiration without flushing? SUMMARY ANSWER: Follicular flushing with the modified flushing system did not increase the number of oocytes, but increased the procedure duration. WHAT IS KNOWN ALREADY: Most studies on follicular flushing were performed with conventional double-lumen needles in patients who were normal responders...
April 1, 2017: Human Reproduction
https://www.readbyqxmd.com/read/28331549/dynamics-of-5-methylcytosine-and-5-hydroxymethylcytosine-during-pronuclear-development-in-equine-zygotes-produced-by-icsi
#17
Sonia Heras, Katrien Smits, Catharina De Schauwer, Ann Van Soom
BACKGROUND: Global epigenetic reprogramming is considered to be essential during embryo development to establish totipotency. In the classic model first described in the mouse, the genome-wide DNA demethylation is asymmetric between the paternal and the maternal genome. The paternal genome undergoes ten-eleven translocation (TET)-mediated active DNA demethylation, which is completed before the end of the first cell cycle. Since TET enzymes oxidize 5-methylcytosine to 5-hydroxymethylcytosine, the latter is postulated to be an intermediate stage toward DNA demethylation...
2017: Epigenetics & Chromatin
https://www.readbyqxmd.com/read/28331070/cytoplasmic-flows-as-signatures-for-the-mechanics-of-mitotic-positioning
#18
Ehssan Nazockdast, Abtin Rahimian, Daniel Needleman, Michael Shelley
The proper positioning of mitotic spindle in the single-cell Caenorhabditis elegans embryo is achieved initially by the migration and rotation of the pronuclear complex (PNC) and its two associated astral microtubules (MTs). Pronuclear migration produces global cytoplasmic flows that couple the mechanics of all microtubules, the PNC, and the cell periphery with each other through their hydrodynamic interactions (HIs). We present the first computational study that explicitly accounts for detailed HIs between the cytoskeletal components and demonstrate the key consequences of HIs on the mechanics of pronuclear migration...
March 22, 2017: Molecular Biology of the Cell
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
#19
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
#20
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...
2017: Cellular Physiology and Biochemistry
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