Engineered reproductive isolation in animals
AssigneeThe Regents of the University of California
InventorsOmar Akbari, Anna Buchman
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Abstract
Provided herein are methods of engineering a reproductive barrier in an insect including (a) introducing into a first insect a first nucleic acid sequence, where the first nucleic acid sequence targets a genomic sequence, where the genomic sequence is proximal to a transcription start site of a gene product; and (b) introducing into a second insect a second nucleic acid sequence encoding a nuclease-deficient endonuclease; and (c) genetically crossing the first insect with the second insect, where the second insect includes a second copy of the genomic sequence where the second copy of the genomic sequence is proximal to a transcription start site of a second copy of the gene product.
Claims45
A method of engineering a reproductive barrier in an insect comprising:
- (a) introducing into a first insect
- a first nucleic acid sequence, wherein the first nucleic acid sequence targets a genomic sequence, wherein the genomic sequence is proximal to a transcription start site of a gene product;
- (b) introducing into a second insect
- a second nucleic acid sequence encoding a nuclease-deficient endonuclease;
- and
- (c) genetically crossing the first insect with the second insect, wherein the second insect comprises a second copy of the genomic sequence wherein the second copy of the genomic sequence is proximal to a transcription start site of a second copy of the gene product,
- wherein the first nucleic acid sequence targets the genomic sequence and/or the second copy of the genomic sequence, and
- wherein in the progeny of the genetic cross between the first insect and the second insect the nuclease-deficient endonuclease binds to the genomic sequence and/or the second copy of the genomic sequence, thereby promoting expression of the (i) gene product and/or (ii) the second copy of the gene product.
The method of any one of claims 27-32, wherein the endonuclease comprises a Streptococcus pyogenes Cas9 (SpCas9), a Staphylococcus aureus Cas9 (SaCas9), a Francisella novicida Cas9 (FnCas9), or a variant thereof.
A method of engineering a reproductive barrier in an insect comprising:
- (a) introducing into a first insect
- a first nucleic acid sequence, wherein the first nucleic acid sequence targets a genomic sequence that is proximal to a transcription start site of a gene product;
- and
- an endonuclease, wherein the endonuclease cleaves the genomic sequence, thereby creating a mutation in the genomic sequence;
- and
- (b) introducing into a second insect
- a second nucleic acid sequence encoding a nuclease-deficient endonuclease;
- and
- (c) genetically crossing the first insect with the second insect, wherein the second insect comprises a second copy of the genomic sequence, that is proximal to a transcription start site of a second copy of the gene product,
- wherein the first nucleic acid sequence targets the second copy of the genomic sequence, and
- wherein in the progeny of the genetic cross between the first insect and the second insect the nuclease-deficient endonuclease binds to the second copy of the genomic sequence, thereby promoting expression of the second copy of the gene product.
Dependencies are inferred from the claim text.