The disclosure provides, inter alia, methods of improving sperm function and related methods of fertilization, together with preparations of activated or potentiated sperm. The methods provided by the disclosure, in some embodiments entail energy depletion with subsequent staged reintroduction of different energy sources. The disclosure additionally provides articles of manufacture suitable for performing the methods provided by the invention. The invention provides kits for separating sperm and for processing and preparing sperm for, in some embodiments, IVF or IUI. Also provided are nutrient free reagents useful preparing sperm.

The disclosure provides, inter alia, methods of improving sperm function and related methods of fertilization, together with preparations of activated or potentiated sperm. The methods provided by the disclosure, in some embodiments entail energy depletion with subsequent staged reintroduction of different energy sources. The disclosure additionally provides articles of manufacture suitable for performing the methods provided by the invention. The invention provides kits for separating sperm and for processing and preparing sperm for, in some embodiments, IVF or IUI. Also provided are nutrient free reagents useful preparing sperm.

A dorsal aortic injection method of chicken embryos for improving transgenic efficiency, comprising the following steps: injecting adeno-associated virus particles into the dorsal aorta of each 2.5-day-old chicken embryo outside eggshell, individually transferring the injected chicken embryos into their recipient eggshells (coming from double yolk eggs), and proceeding to hatch. The disclosure realizes the outside-eggshell injection of the chicken embryo dorsal aorta through the modified dorsal aortic injection method, improves the accuracy of injection, and also realizes the direct observation of the development process of the chicken embryos. The survival rate of embryos, hatch rate and the efficiency of transgene expression in the modified dorsal aortic injection group are significantly higher than those in the subgerminal cavity injection group and the classic dorsal aortic injection group, indicating that the modified method improves the hatch rate of transgenic chicken embryos and EGFP expression efficiency.

The present invention provides modified oocytes having a nuclear genome derived from a first oocyte and cytoplasm derived from a second oocyte from a different subject, and methods for making and using such modified oocytes. The methods and compositions of the present invention can be useful in a variety of settings including, but not limited to, in in vitro fertilization (“IVF”) procedures.

The present invention provides modified oocytes having a nuclear genome derived from a first oocyte and cytoplasm derived from a second oocyte from a different subject, and methods for making and using such modified oocytes. The methods and compositions of the present invention can be useful in a variety of settings including, but not limited to, in in vitro fertilization (“IVF”) procedures.

Methods for gene editing of embryos in vitro are provided. The methods typically include contacting an embryo in vitro with an effective amount of non-enzymatic (e.g., non-nuclease) gene editing active agent(s) optionally encapsulated, entrapped, complexed to or dispersed in polymeric particles to induce at least one alteration in the genome of the embryo. The embryo can be a single cell zygote, however, treatment of male and female gametes prior to fertilization, and embryos having 2, 4, 8, or 16 cells, and including not only zygotes, but also morulas and blastocysts are also provided. Typically, the embryo is contacted with the particles on culture days 0-6 during or following in vitro fertilization.

Methods for gene editing of embryos in vitro are provided. The methods typically include contacting an embryo in vitro with an effective amount of non-enzymatic (e.g., non-nuclease) gene editing active agent(s) optionally encapsulated, entrapped, complexed to or dispersed in polymeric particles to induce at least one alteration in the genome of the embryo. The embryo can be a single cell zygote, however, treatment of male and female gametes prior to fertilization, and embryos having 2, 4, 8, or 16 cells, and including not only zygotes, but also morulas and blastocysts are also provided. Typically, the embryo is contacted with the particles on culture days 0-6 during or following in vitro fertilization.

The present invention relates to methods for transfecting cells. In particular, the present invention relates to methods of transfecting primordial germ cells in avians, and to methods of breeding avians with modified traits.

The present invention relates to methods for transfecting cells. In particular, the present invention relates to methods of transfecting primordial germ cells in avians, and to methods of breeding avians with modified traits.

Factors for extending the ability of isolated pluripotent stem cells to generate extraembryonic lineages in vivo, following in vitro culture, herein, chemical extenders of pluripotency (CEP). Methods of extending the ability of a pluripotent cell to generate embryonic and extraembryonic lineages. The cell to be reprogrammed is contacted with effective amounts of the CEPs for a sufficient period of time to reprogram the cell into a chemically induced extended pluripotent cell (ciEPSC). ciEPSC are identified as an extended pluripotent cell based on properties including: (i) morphologically and (ii) functionally for example, based on their ability contribute to both TE and ICM, in vivo. The ciEPSCs can be cultured or induced to differentiate into cells of a desired type, and used in a number of applications, including but not limited to cell therapy and tissue engineering.