US5523222A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Polyelectrolyte DNA conjugation and genetic transformation of an animal
Assignee: Virginia Tech Intellectual Properties Inc
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Filing: 1994-08-05
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Granted: 1996-06-04
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Expires: 2014-08-05
Abstract
Abstract The present invention provides a method of obtaining an organism which has been characterized as having cells containing exogenous genetic material which includes any sequence of DNA that can be distinguished as exogenous by known molecular biological analysis by insertion of genetic material into an animal's genetic makeup. The insertion of the genetic material is done by inserting DNA that has been complexed with molecules that allow the DNA to be inserted into the chromosomes when injected into the cytoplasm, perivitelline space, or placed in surrounding culture media to be taken up and incorporated into the genome. When the DNA is complexed into the polyelectrolyte molecules by electrostatic attraction, the electric charge of DNA of the complex is partially to substantially neutralized. The present method does not require the genetic material to be introduced into the embryo at a particular stage in development.
Inventors
- 👤 Raymond L. Page
- 👤 William H. Velander
- 👤 John L. Johnson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 17:12
CPC Classifications
A; A01; A01K; A01K67/00; A01K67/027; A01K67/0275; C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; C12N15/89; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6464; C12Y; C12Y304/00; C12Y304/21; C12Y304/21069; A01K2217/00; A01K2217/05
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
117 words
A method for inserting a DNA segment into the genome of a recipient animal selected from the group consisting of mammals and birds, comprising: forming an electrostatic complex of a polycation and a DNA segment, wherein the polycation is present in the complex in an amount effective to neutralize the negative electric charges of the DNA segment to a degree sufficient to allow insertion of the DNA segment into a chromosome of the recipient animal, injecting the polycation/DNA complex in vitro into the cytoplasm of an animal cell selected from the group consisting of an unfertilized ovum and a one-celled embryo, and testing the resulting animal to verify the insertion of the DNA segment into its genome.
Claim 2
Independent Claim
117 words
A method for inserting a DNA segment into the genome of a recipient animal selected from the group consisting of mammals and birds, comprising: forming an electrostatic complex of a polycation and a DNA segment, wherein the polycation is present in the complex in an amount effective to neutralize the negative electric charges of the DNA segment to a degree sufficient to allow insertion of the DNA segment into a chromosome of the recipient animal, injecting the polycation/DNA complex in vitro into the cytoplasm of an animal cell selected from the group consisting of an unfertilized ovum and a one-celled embryo, and testing the resulting animal to verify the insertion of the DNA segment into its genome.
Claim 3
Dependent Claim
11 words
A method according to claim 1, wherein the animal a mammal.
Claim 4
Dependent Claim
20 words
A method according to claim 2, wherein the animal is selected from the group consisting of mice, pigs and cattle.
Claim 5
Dependent Claim
13 words
A method according to claim 1, wherein said cell is an unfertilized ovum.
Claim 6
Dependent Claim
13 words
A method according to claim 1, wherein said cell is a one-cell embryo.
Claim 7
Dependent Claim
24 words
A method according to claim 1, wherein the inserted polycation/DNA complex has a polycation/DNA molar charge ratio of from about 5:1 to about 0.25:1.
Claim 8
Dependent Claim
22 words
A method according to claim 6, wherein the molar charge ratio of the polycation/DNA complex is from about 2:1 to about 1:1.
Claim 9
Dependent Claim
22 words
A method according to claim 7, wherein the polycation is polylysine and the molar charge ratio of the complex is about 1:1.
Claim 10
Dependent Claim
16 words
A method according to claim 1, wherein the electrostatic complex consists essentially of polycation and DNA.
Claim 11
Dependent Claim
12 words
A method according to claim 1, wherein said animal is a bird.
Claim 12
Dependent Claim
12 words
A method according to claim 10, wherein said bird is a chicken.
Claim 13
Dependent Claim
12 words
A method according to claim 3, wherein said animal is a pig.
Claim 14
Dependent Claim
11 words
A method according to claim 2, wherein said animal is bovine.
Claim 15
Dependent Claim
12 words
A method according to claim 3, wherein said animal is a mouse.
References Cited (Backward Citations)
US4873191A
Cited
US5166320A
Cited
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