Artificial Uterus and Artificial Embryos: Unsolved Tasks.

Ex utero embryo culture is one of the important tasks that has not been completely solved. In the first part of the current review, we focus on potential roles ex utero mammalian embryo culture could play, such as in infertility treatment or ex utero embryo treatment. Particularly, we discuss ex utero genetic therapy of embryos for correction and prevention of congenital pathologies.

Biological effects of femtosecond and millisecond lasers application for assisted hatching in mouse embryos.

Purpose: Laser-assisted hatching (LAH) is a typical procedure in ART. It is used when embryos are unable to hatch. To reduce the potential damage to embryos caused by millisecond laser typically used to create zona pellucida openings, we have developed a protocol for femtosecond laser microsurgery.

Mutations in Filamin C Associated with Both Alleles Do Not Affect the Functioning of Mice Cardiac Muscles.

Filamin C (FLNC) is a structural protein of muscle fibers. Mutations in the gene are known to cause myopathies and cardiomyopathies in humans. Here we report the generation by a CRISPR/Cas9 editing system injected into zygote pronuclei of two mouse strains carrying filamin C mutations-one of them (AGA) has a deletion of three nucleotides at position c.

E12.5 whole mouse embryo culture.

Ex utero culture of postimplantation embryos remains one of the unsolved tasks in developmental biology. This technique may be required for infertility treatment, observation of embryo development and assessing the embryotoxicity of certain chemical agents. We describe novel method for E12.

Genetically modified mice as a tool for the study of human diseases.

Modeling a human disease is an essential part of biomedical research. The recent advances in the field of molecular genetics made it possible to obtain genetically modified animals for the study of various diseases. Not only monogenic disorders but also chromosomal and multifactorial disorders can be mimicked in lab animals due to genetic modification.

Genetically Engineered Mice Unveil In Vivo Roles of the Mediator Complex.

The Mediator complex is a multi-subunit protein complex which plays a significant role in the regulation of eukaryotic gene transcription. It provides a platform for the interaction of transcriptional factors and RNA polymerase II, thus coupling external and internal stimuli with transcriptional programs. Molecular mechanisms underlying Mediator functioning are intensively studied, although most often using simple models such as tumor cell lines and yeast.

Limitations of Tamoxifen Application for In Vivo Genome Editing Using Cre/ER System.

Inducible Cre-dependent systems are frequently used to produce both conditional knockouts and transgenic mice with regulated expression of the gene of interest. Induction can be achieved by doxycycline-dependent transcription of the wild type gene or OH-tamoxifen-dependent nuclear translocation of the chimeric Cre/ER protein. However, both of these activation strategies have some limitations.

AAV infection of bovine embryos: Novel, simple and effective tool for genome editing.

Adeno-associated viruses (AAV) are widely used in the field of genetically modified organism production. In this work, transduction of bovine embryos by AAV was selected as a potential approach to perform genetic modifications: we have used recombinant AAV to produce GFP-positive bovine embryos. Five different AAV serotypes were used to evaluate their ability to deliver genetic material into the bovine embryos.

Controlled hatching at the prescribed site using femtosecond laser for zona pellucida drilling at the early blastocyst stage.

Purpose: To study whether the application of femtosecond laser pulses for zona pellucida (ZP) drilling of blastocysts at the embryonic or abembryonic poles can promote hatching to start immediately through the hole formed and ensure high hatching rates and embryo viability.

Methods: Mouse blastocyst (E3.5) ZP were microdissected with femtosecond laser pulses (514-nm wavelength, 280-fs pulse duration, 2.

The Analysis of Quality of Ovarian Follicle Culture Systems Using Time-Lapse Microscopy and Quantitative Real-Time PCR.

Background: The aim of ovarian follicle culture is to obtain mature oocytes. To evaluate the efficiency of culture system, the status of the cultured oocyte can be analyzed.

Methods: The preantral ovarian follicles retrieved from 14-day-old C57Bl/6J mice were cultured in 3D alginate hydrogel.

Application of femtosecond laser microsurgery in assisted reproductive technologies for preimplantation embryo tagging.

Femtosecond laser pulses were applied for precise alphanumeric code engraving on the (ZP) of mouse zygotes for individual embryo marking and their identification. The optimal range of laser pulse energies required for safe ZP microsurgery has been determined. ZP was marked with codes in three different planes to simplify the process of embryo identification.

Femtosecond laser is effective tool for zona pellucida engraving and tagging of preimplantation mammalian embryos.

Purpose: Our purpose was to study whether application of femtosecond laser pulses for alphanumeric code marking in the volume of zona pellucida (ZP) could be effective and reliable approach for direct tagging of preimplantation embryos.

Methods: Femtosecond laser pulses (wavelength of 514 nm, pulse duration of 280 fs, repetition rate of 2.5 kHz, pulse energy of 20 nJ) were applied for precise alphanumeric code engraving on the ZP of mouse embryos at the zygote stage for individual embryo marking and their accurate identification.