摘要:
The present invention provides a microinjection assembly including a microscope, a microinjection system comprising a micromanipulator, a micropipette and a piezo-electric oscillator, and an obliquely angled macro monitoring unit. The present invention also provides methods of microinjecting the germinal disk of an avian egg, thereby delivering a transgenic nucleus, spermatozoon or isolated nucleic acid to the avian embryo. The avian ovum may be returned to a female bird for hard-shell deposit and laying of the egg for hatching as a transfected bird.
摘要:
The present invention relates generally to novel methods of producing transgenic chickens that generate antibodies or immunoglobulin polypeptides in whites of eggs. More specifically, one embodiment of the present invention relates to methods of inserting immunoglobulin-encoding transgenes into avian sperm cells for transfer to ova to generate transgenic zygotes. The transgenes may include at least two immunoglobulin-encoding nucleic acid sequences and an internal ribosome entry site (IRES) that allow the immunoglobulin polypeptides to be expressed by chicken cells and hence in egg whites.
摘要:
The invention provides methods for integrating a heterologous polynucleotide into the genome of an avian cell. The methods deliver to an avian cell a polynucleotide and a source of integrase activity that mediates recombination between the polynucleotide and the genomic DNA of the avian cell. The invention provides modified avian or artificial chromosomes as vectors to shuttle transgenes or gene clusters into an avian genome. Another aspect of the invention are avian cells genetically modified with a transgene vector. One cell line for the delivery and integration of a transgene comprises a heterologous attP site and, optionally, a region for expressing the integrase. Methods are also included for the production of a heterologous polypeptide by transgenic avian tissue involve integrating a heterologous polynucleotide into the avian genome. The present invention also relates to methods of producing transgenic chickens which include introducing into an avian cell a nucleic acid comprising a transgene and an integrase activity in addition to a cationic polymer and/or a nuclear localization signal and introducing the avian cell into a recipient avian wherein the recipient avian produces an offspring which includes the transgene. Also included are methods of dispersing a nucleic acid in a cell.
摘要:
This invention includes methods for treating conditions with pharmaceutical compositions that comprise glycosylated interferon alpha. Pharmaceutical compositions that can be employed in the present invention include glycosylated interferon alpha (2) species in combination with pharmaceutical carriers, pharmaceutical excipients and/or other agents.
摘要:
This invention provides vectors and methods for the stable introduction of exogenous nucleic acid sequences into the genome of avians in order to express the exogenous sequences to alter the phenotype of the avians or to produce desired proteins. In particular, transgenic avians are produced which express exogenous sequences in their oviducts and which deposit exogenous proteins into their eggs. Avian eggs that contain exogenous proteins are encompassed by this invention. The instant invention further provides novel forms of interferon and erythropoietin which are efficiently expressed in the oviduct of transgenic avians and deposited into avian eggs.
摘要:
The invention provides methods for integrating a heterologous polynucleotide into the genome of an avian cell. The methods deliver to an avian cell a polynucleotide and a source of integrase activity that mediates recombination between the polynucleotides and the genomic DNA of the avian cell. The invention provides modified avian or artificial chromosomes as vectors to shuttle transgenes or gene clusters into an avian genome. Another aspect of the invention are avian cells genetically modified with a transgene vector. One cell line for the delivery and integration of a transgene comprises a heterologous attP site and, optionally, a region for expressing the integrase. Methods are also included for the production of a heterologous polypeptide by transgenic avian tissue involve integrating a heterologous polynucleotide into the avian genome. Subsequently, a mature transgenic avian is derived by transferring the transgenic blastodermal cells to an embryo and allowing that embryo to develop fully.
摘要:
Methods for producing cloned non-human animals by nuclear transfer and by combinations of nuclear transfer and ovum transfer, employing near-infrared visualization of the recipient cell nucleus are described herein. Transgenic, knock-out, and knock-in avians are provided. Method for producing eggs which contain exogenous proteins are encompassed by the present invention. Thus, the instant invention satisfies the need for an effective route to the generation of cloned avians.
摘要:
The invention includes transchromosomal avians and transchromosomal avian cells and methods for the introduction of artificial chromosomes into the genome of avians and avian cells.
摘要:
The present invention provides isolated and recombinant avian nucleic acid molecules comprising at least one avian MAR and an avian nucleic acid sequence encoding an ovalbumin transcriptional regulatory region. The isolated nucleic acid of the present invention is useful for reducing chromosomal positional effects upon the transcription of a transgene operably linked to the ovalbumin transcriptional regulatory region and transfected into a recipient avian cell The recombinant nucleic cid molecules of the present invention may further comprise a polyadenylation signal sequence or an avian 3' domain, and optionally, an internal ribosome entry site for expression of an operably linked heterologous nucleic acid insert in a transfected avian cell.
摘要:
The present invention relates generally to novel methods of producing transgenic chickens that generate antibodies or immunoglobulin polypeptides in whites of eggs. More specifically, one embodiment of the present invention relates to methods of inserting immunoglobulin-encoding transgenes into avian sperm cells for transfer to ova to generate transgenic zygotes. The transgenes may include at least two immunoglobulin-encoding nucleic acid sequences and an internal ribosome entry site (IRES) that allow the immunoglobulin polypeptides to be expressed by chicken cells and hence in egg whites.