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The use of engineered nucleases in one-cell stage mouse embryos is emerging as an efficient alternative to conventional gene targeting in mouse embryonic stem (ES) cells.
Intact, manually separated blastomeres from wild type and maternal knockout E-cadherin+/− two- to eight-cell stage mouse embryos were used.
Based on recent advances, here we provide experimentally validated, detailed guidelines for generating non-homologous end-joining (NHEJ -mediated mutaNHEJ -mediatedroinjecting TALENs and RGENs into the cytoplasmutanthe pronucleus of one-cell stage micee embyyos.
Objective: To examine the rescue of mouse embryos from 2-cell blocks by the microinjection of maturation promoting factor (MPF) extracted from matured Xenopus eggs into one of the blastomeres of 2-cell stage mouse embryos.
Cells were derived from E13.5 stage mouse embryos.
2-cell stage mouse embryos were injected in both blastomeres with a tubulin-GFP mRNA.
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As a test, the researchers injected a solution of Kit microRNAs into otherwise wild-type one-cell-stage mouse embryos.
To gain insights into the molecular determinants of foregut development, we performed microarray analyses to identify endoderm-enriched transcripts in the foregut of early-somite-stage mouse embryos.
Using Ca2+ imaging and patch-clamp techniques, we studied requirements for generation of spontaneous rhythmic action potentials (APs) in late-stage mouse ESCs.
To test this hypothesis, the researchers tagged the reprogrammed cells, called induced pluripotent stem (iPS) cells, with a fluorescent dye and injected them into early-stage mouse embryos.
For example, when introduced into early-stage mouse embryos, they produced chimeric animals that have body tissues containing two different genomes, the team reports today in the journal Cell Stem Cell.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com