Method for inducing targeted meiotic recombinations
AssigneesMeiogenix, Institut Curie, Centre National De La Recherche Scientifique, Universite Pierre Et Marie Curie (Paris 6)
InventorsGIACOMO BASTIANELLI, ALEXANDRE SERERO, ALAIN NICOLAS
Life of this patent
Estimating the term…
Abstract
The present invention relates to a fusion protein comprising a Cas9 domain and a Spo11 domain, as well as the use of this protein to induce targeted meiotic recombinations in a eukaryotic cell.
Claims22
1-27. (canceled)
A method for inducing targeted meiotic recombinations in a eukaryotic cell comprising:
- introducing into said cell:
- a) a fusion protein comprising a Cas9 domain and a Spo11 domain, or a nucleic acid encoding said fusion protein;
- and
- b) one or more guide RNAs or one or more nucleic acids encoding said guide RNAs, said guide RNAs comprising an RNA structure for binding to the Cas9 domain of the fusion protein and a sequence complementary to the targeted chromosomal region;
- and
- inducing said cell to enter meiotic prophase I.
A fusion protein comprising a Cas9 domain and a Spo11 domain.
A method for generating variants of a non-human eukaryotic organism comprising introducing into a cell of said non-human eukaryotic organism:
- a) a fusion protein comprising a Cas9 domain and a Spo11 domain, or a nucleic acid encoding said fusion protein;
- and
- b) one or more guide RNAs, or one or more nucleic acids encoding said guide RNAs, said guide RNAs comprising an RNA structure for binding to the Cas9 domain and a sequence complementary to a targeted chromosomal region;
- and
- inducing said cell to enter meiotic prophase I;
- obtaining a cell or cells having the desired recombination(s) at the targeted chromosomal region(s);
- and
- generating a variant of the organism from said recombinant cell.
A method for identifying or locating the genetic information encoding a characteristic of interest in a eukaryotic cell genome comprising:
- introducing into the eukaryotic cell:
- a) a fusion protein comprising a Cas9 domain and a Spo11 domain, or a nucleic acid encoding said fusion protein;
- and
- b) one or more guide RNAs, or one or more nucleic acids encoding said guide RNAs, said guide RNAs comprising an RNA structure for binding to the Cas9 domain and a sequence complementary to a targeted chromosomal region;
- and
- inducing said cell to enter meiotic prophase I;
- obtaining a cell or cells having the desired recombination(s) at the targeted chromosomal region(s);
- and
- analyzing the genotypes and phenotypes of the recombinant cells in order to identify or to locate the genetic information encoding the characteristic of interest.
Dependencies are inferred from the claim text.