Materials and methods for producing animals with short hair
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Publication Number
US-10716298-B2
Publication Date
2020-07-21
Expiration Date
Abstract
The subject invention provides materials and methods for producing animals with short hair length. In a preferred embodiment, this is accomplished by altering in the animal the nucleotide sequence that encodes the prolactin receptor (PRLR) protein such that a truncated version of the protein is produced. Advantageously, and surprisingly, the truncated protein produced according to the subject invention retains lactogenic functionality, but causes the animal to have a short-hair coat.
Core Innovation
The invention relates to producing animals with short hair (slick phenotype) by altering the prolactin receptor (PRLR) genomic nucleotide sequence to express a truncated PRLR protein. The altered PRLR is described as retaining lactogenic functionality while causing the short hair phenotype, with the truncated PRLR protein lacking conserved C-terminal residues from the full-length wild-type PRLR.
The document describes genetically engineered cattle/bovids in which a truncated PRLR polynucleotide is incorporated into the genome to produce a truncated PRLR protein. A specific truncation embodiment is described in which the truncated PRLR corresponds to a minimal lactogenic fragment and excludes the amino-acid region corresponding to the deleted/absent C-terminal portion of the wild-type PRLR.
The document further identifies a mutation (A461VfsX1) and provides a rationale that deleted conserved C-terminal residues are non-essential for milk production, while enabling the slick phenotype. Additional disclosure themes include sequence relationships and characterization or detection of PRLR truncation mutants using antibody-based and analytical approaches.
Claims Coverage
The partial content identifies one independent claim and several dependent claims. The independent claim is directed to a modified bovine cell with a genome-incorporated polynucleotide encoding a truncated PRLR protein of a defined length, with dependent claims refining the introduced polynucleotide identity relative to a defined SEQ ID, specifying the truncated amino-acid segment, and limiting the genome-engineering approach.
Genome-incorporated truncated PRLR polynucleotide in a modified bovine cell
The method includes obtaining a bovine cell having genomic DNA comprising a full-length wild-type PRLR protein, and introducing a polynucleotide encoding a truncated PRLR protein into the bovine cell such that the polynucleotide is incorporated into the genome to produce the modified bovine cell, wherein the truncated PRLR protein is 426 amino acids in length.
Sequence identity of the introduced truncated PRLR polynucleotide to SEQ ID NO: 3
The polynucleotide encoding the truncated PRLR protein comprises a sequence having at least 90% sequence identity to the sequence of SEQ ID NO: 3.
Defined truncated PRLR amino-acid segment composition
The truncated PRLR protein consists of amino acids 1-426 of SEQ ID NO.: 7.
Tightened sequence identity of the introduced truncated PRLR polynucleotide to SEQ ID NO: 3
The polynucleotide encoding the truncated PRLR protein comprises a sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 3.
CRISPR/Cas or TALEN-mediated genome engineering to introduce the truncated PRLR polynucleotide
The polynucleotide encoding the truncated PRLR protein is introduced and incorporated into the bovine cell genome using a CRISPR/Cas system or TALEN-mediated genome engineering.
Overall claim coverage centers on producing a modified bovine cell by incorporating a polynucleotide encoding a truncated PRLR protein of defined length (426 amino acids), with refinements specifying sequence-identity thresholds relative to SEQ ID NO: 3, defining the truncated PRLR amino-acid segment (amino acids 1-426 of SEQ ID NO: 7), and enabling incorporation via CRISPR/Cas or TALEN genome engineering.
Stated Advantages
The truncated PRLR protein is described as retaining lactogenic functionality while producing the short hair (slick phenotype).
Deleted conserved C-terminal residues are described as non-essential for milk production.
Documented Applications
Producing animals with short hair (slick phenotype) by engineering PRLR genomic nucleotide sequence to express a truncated PRLR protein.
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