Materials and methods for preventing transmission of a particular chromosome
AssigneeAggenetics, Inc.
InventorJames WEST
Life of this patent
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Abstract
Provided herein are material and methods for changing gene expression in select sex chromosomes. The materials and methods of the subject invention can be used to produce non-human transgenic animals that produce progeny of a predetermined gender and to generate non-human transgenic animals that produce single-sexed semen.
Claims37
1-2. (canceled)
A method for preventing or forcing transmission of a particular chromosome in a male non-human transgenic mammal, comprising:
- providing a genetic construct comprising an exogenous nucleic acid sequence operably linked to a promoter that activates expression of the exogenous nucleic acid sequence post-meiotically in a developing sperm cell;
- wherein
- (A) the exogenous nucleic acid sequence comprises an untranslated region (UTR) that tethers a transcript transcribed from the nucleic acid sequence to a cytoskeletal structure of the sperm cell;
- and the exogenous nucleic acid sequence encodes at least one protein that prevents transmission by inhibiting progressivity, motility, and/or penetration ability of a sperm cell or induces sperm cell death or forces transmission by promoting progressivity, motility, and/or penetration ability of a sperm cell or inhibits sperm cell death, and the at least one protein optionally comprises a membrane-association sequence that tethers to a cytoskeletal structure of the sperm cell;
- or
- (b) wherein the exogenous nucleic acid sequence comprises (i) a short hairpin RNA (shRNA) or (ii) one small interfering RNA (siRNA) inserted into a micro RNA (miR) cassette, operably linked to an untranslated region (UTR) that tethers a transcript transcribed from the nucleic acid sequence to a cytoskeletal structure of the sperm cell, wherein the shRNA or the siRNA inserted into miR cassette prevents transmission by inhibiting expression of at least one gene involved in survival, motility, progressivity and/or penetration ability of the sperm cell or forces transmission by inhibiting at least endogenous one gene expressing a protein involved in survival, motility, progressivity and/or penetration ability of the sperm cell;
- and wherein the genetic construct further comprises a second exogenous nucleic acid sequence encoding the protein, and wherein for forcing transmission the second exogenous nucleic acid sequence encoding comprises third base wobbles such that the shRNA or siRNA inserted into miR cassette not inhibit expression of the second exogenous nucleic acid sequence encoding the protein;
- and introducing the construct into the particular chromosome in a cell of the non-human animal.
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28-29. (canceled)
A genetic construct comprising a nucleic acid sequence operably linked to a promoter that activates expression of the nucleic acid sequence post-meiotically in a developing sperm cell, wherein the nucleic acid sequence comprising an untranslated region (UTR) that tethers a transcript transcribed from the nucleic acid sequence to a cytoskeletal structure of a sperm cell;
- and
- wherein the nucleic acid sequence encodes a dominant-negative Slc26a8 protein or at least one protein that inhibits progressivity, motility, and/or penetration ability of a sperm cell or induces sperm cell death or encodes at least one protein that promotes progressivity, motility, and/or penetration ability of a sperm cell or inhibits sperm cell death, and the at least one protein optionally comprises a membrane-insertion sequence that tethers to a cytoskeletal structure of the sperm cell.
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36-39. (canceled)
A genetic construct comprising a nucleic acid sequence operably linked to a RNA polymerase III (pol III) promoter, wherein the nucleic acid sequence optionally comprising an untranslated region (UTR) that tethers a transcript transcribed from the nucleic acid sequence to a cytoskeletal structure of a sperm cell;
- and
- wherein the nucleic acid sequence comprises a short hairpin RNA (shRNA) promoting survival, motility, progressivity and/or penetration ability of the sperm cell.
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A nucleic acid molecule comprising at least one small interfering RNA (siRNA) for at least one protein that enables progressivity, motility and/or penetration ability of a sperm cell;
- wherein the at least one siRNA is inserted into a micro RNA (miR) cassette, which miR cassette comprises at least one sequence homologous to a sequence of a 3′UTR region of a gene expressed in late spermatogenesis.
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Dependencies are inferred from the claim text.