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US 12,213,466 B2Granted
Process for using CRISPR to transfect primordial germ cells in avians
AssigneesCommonwealth Scientific and Industrial Research Organisation, Aviagen
InventorScott Geoffrey Tyack
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Abstract
The present invention relates to processes for transfecting cells. In particular, the present invention relates to processes for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC).
Claims8
A method of incorporating exogenous DNA into the genome of chicken or quail primordial germ cells (PGCs), the method comprising:
- injecting a transfection mixture comprising:
- a) a DNA polynucleotide encoding a guide RNA (gRNA) and a Cas9 nuclease;
- b) exogenous DNA for integration into the genome of the PGCs;
- and
- c) a cationic lipid
- into a blood vessel of:
- i) a stage 13-17 chicken embryo, or
- ii) an equivalent stage quail embryo,
- such that the exogenous DNA is integrated into the genome of the PGCs.
6 more elements
Dependencies are inferred from the claim text.