North Carolina State University

The United States of America, As Represented by the Secretary, Department of Health and Human Services

University of Rochester

Embrex, Inc.

James N Petitte

Zengming Yang

Samuel Lloyd Pardue

Catherine A Ricks

Susan D'Costa

Paul E Mozdziak

Sally E Spence

Paul R Reynolds

Stephen Shears

Il-Kuk Chang

Yi Guo Zhang

Yonghong Song

A transgenic animal such as a transgenic snake or other reptile that expresses a heterologous expression product is described, along with methods of making the same. In general, the animal comprises cells containing a sequence encoding the heterologous expression product. The sequence encoding the heterologous expression product is integrated into the genome of the animal (e.g., in some or all cells thereof, and in some embodiments into germ cells thereof). The sequence encoding the heterologous expression product is, in general, operatively associated with an expression sequence or promoter. The animals are useful for, among other things, testing of repellents, testing of toxicological compounds, as teaching aids, for venom production, etc.

Transgenic reptiles

Methods for modulating primordial germ cell (PGC) numbers and/or development in avians are provided. In one embodiment, the presently disclosed subject matter provides a method for modulating primordial germ cells numbers in an avian embryo comprising immunizing a female bird with an antigen associated with primordial germ cells, whereby an egg produced by the female bird comprises a sufficiently high concentration of antibodies specific for the antigen to modulate numbers of endogenous PGCs in an avian embryo present within in the egg. Also provided are methods for producing chimeric avians, methods for increasing the proportion of male birds in a plurality of eggs, methods of producing avian gametes, and methods for enhancing germ line transmission of nucleic acids in birds.

Depletion of endogenous primordial germ cells in avian species

Transgenic snakes and methods of making

A method of producing undifferentiated avian cells expressing an embryonic stem cell phenotype. The method includes the steps of collecting avian gonadal cells including primordial germ cells from an avian embryo after the formation of the primitive streak; depositing the avian gonadal cells in contact with a preconditioned feeder matrix; and growing the avian gonadal cells on the pre-conditioned feeder matrix in the presence of media for a time sufficient to produce an avian cell culture consisting essentially of undifferentiated avian cells expressing an embryonic stem cell phenotype.

Culture of undifferentiated chicken cells

The present invention provides an isolated animal cell comprising an exogenous nucleic acid encoding a mutated phytase, wherein the cell produces phytase and secretes the phytase from the cell. The present invention also provides an animal having a phenotype not naturally occurring, characterized by secretion of phytase into the lumen of the gastrointestinal tract of the animal, the phenotype being conferred by a transgene contained in the cells of the animal, the transgene comprising a nucleic acid sequence encoding phytase and methods of making said animal.

Use of a transgene encoding a vertebrate phytase to increase capacity to utilize phytic acid in livestock feed

The present invention provides methods for the production of avian gametes, comprising reducing endogenous primordial germ cells (PGCs) in a recipient bird in ovo and introducing donor primordial germ cells from another bird into the recipient bird in ovo. The present invention further provides a method of increasing the ratio of male offspring from a female bird, comprising reducing endogenous PGCs in the female bird in ovo, introducing donor PGCs from another bird into the female bird in ovo, and incubating the female bird to hatch. The female bird is raised to sexual maturity and bred to produce a plurality of eggs having a higher proportion of male eggs than in the absence of the inventive methods. In some embodiments, the donor PGCs are from a different species than the recipient bird. In preferred embodiments, endogenous PGCs are reduced using busulfan.

Methods for gamete production in birds

A method of altering the phenotype of a bird comprises introducing a DNA sequence into somatic cells of a bird contained within an egg during in ovo incubation. The DNA sequence is selected to be effective to cause a change in phenotype, such as an increase in growth rate, feed efficiency, or both in the bird after hatch. A DNA sequence may further be selected to increase disease resistance or induce disease prevention by the expression of an antigen over a period of time.

Gene transfer in poultry by introduction of embryo cells in ovo

A method for the production and collection of avian sperm comprises the steps of: providing primordial germ cells from a donor avian species; administering the primordial germ cells to a recipient avian species in ovo; incubating the recipient avian species to hatch; and then collecting sperm of the donor avian species from the recipient avian species. For example, the donor avian species may be a whooping crane, and the recipient avian species may be a sand hill crane. In another example, the donor avian species may be a turkey, and the recipient avian species may be a chicken.

A method of producing undifferentiated avian cells expressing an embryonic stem cell phenotype. The method includes the steps of collecting avian gonadal cells comprising primordial germ cells from an avian embryo after the formation of the primitive streak; depositing the avian gonadal cells in contact with a preconditioned feeder matrix; and growing the avian gonadal cells on the pre-conditioned feeder matrix in the presence of media for a time sufficient to produce an avian cell culture consisting essentially of undifferentiated avian cells expressing an embryonic stem cell phenotype.

Method of producing an undifferentiated avian cell culture using avian primordial germ cells

A method of producing undifferentiated avian cells expressing an embryonic stem cell phenotype is disclosed. The method comprises collecting avian cells from an avian blastoderm prior to formation of the primitive streak, then depositing the avian cells in contact with a mouse fibroblast feeder cell layer, and then growing the avian cells on the mouse fibroblast feeder cell layer in the presence of a media containing leukemia inhibitory factor in a differentiation-inhibiting amount for a time sufficient to produce a sustained avian cell culture. Cell cultures produced by the aforesaid process and veterinary pharmaceutical formulations containing such cells are also disclosed.

Veterinary pharmaceutical formulation containing avian embryonic stem

A method has been developed for the injection of muscle tissue of an embryonic chicken in ovo. An apparatus for such injection in a plurality of eggs simultaneously has also been developed, where the apparatus is composed of a an engaging means for the eggs, an injection means to insert an elongate needle through the eggshell to a depth of about 7/8 inch to 1.5 inch into the shoulder or breast of the embryo, and a positioning means to position the needle about 1 to 5 degrees of offset of the long axis of the egg. The apparatus can delivery a variety of substances to the muscle tissue of the embryonic chicken in ovo.

Growing community of inventors

Raleigh, NC, United States of America

James N Petitte