Claims for Patent: 7,205,148
✉ Email this page to a colleague
Summary for Patent: 7,205,148
Title: | Genome mutation by intron insertion into an embryonic stem cell genome |
Abstract: | Methods of creating mutations in genomic exons by inserting introns into the genomic exons via homologous recombination. Also, methods are provided for introducing modifications into genomic exons by inserting introns into the genomic exons via homologous recombination such that a mature mRNA transcript produced from a genomic region of the genome comprising the genomic exon does not contain the modification are provided. The methods provide for a rapid method for introducing mutations and/or modifications of any type into a mammalian cell genome. |
Inventor(s): | Economides; Aris N. (Tarrytown, NY), Valenzuela; David M. (Yorktown Heights, NY), Davis; Samuel (New York, NY), Yancopoulos; George (Yorktown Heights, NY) |
Assignee: | Regeneron Pharmaceuticals, Inc. (Tarrytown, NY) |
Application Number: | 10/865,424 |
Patent Claims: | 1. An in vitro method of creating a mutation in a genomic exon in a genome of a mouse embryonic stem (ES) cell, comprising: a) constructing a targeting vector
comprising (i) a 5' homology arm, (ii) an intron, wherein the intron comprising a drug selection cassette, wherein the drug selection cassette consists of a drug resistance gene operably linked to a promoter and flanked by a first and a second
site-specific recombination sites, and (iii) a 3' homology arm, wherein the 5' and 3' homology arms contain sequences homologous to corresponding genomic sequences; and b) introducing the targeting vector into the ES cell such that the exon is mutated
by the introduction of the intron.
2. The method of claim 1, wherein the first and the second site-specific recombination sites are same and oriented in the same direction with respect to one another such that excision of DNA sequence between them occurs in the presence of a suitable recombinase. 3. The method of claim 1, wherein the mutation is one or both of (i) an insertion, deletion, or substitution of one ore more nucleotides and (ii) insertion of one or more exogenous DNA sequences. 4. The method of claim 1, wherein the targeting vector further comprises a DNA cassette between the (i) 5' homology arm and (ii) intron, wherein the DNA cassette consists of a third site-specific recombination site, a splice acceptor sequence, and a DNA sequence homologous to a portion of the exon; and the intron further comprises a fourth site-specific recombination site. 5. The method of claim 4, wherein the DNA sequence comprises a mutation. 6. The method of claim 1, wherein the intron further comprises a DNA cassette upstream of the drug selection cassette, wherein the DNA cassette comprises, from 5' to 3' in order, a third site-specific recombination site, a transcription termination sequence, a splice acceptor sequence, and a fourth site-specific recombination site, wherein the transcription termination sequence and the splice acceptor sequence are in antisense orientation, and wherein the third and the fourth site-specific recombination sites are positioned in opposite orientation with respect to one another. 7. The method of claim 6, wherein the third and the fourth site-specific recombination sites are mutated such that the recombination between them cannot occur more than once. 8. The method of claim 6, wherein the DNA cassette further comprises an open reading frame between the transcription termination sequence and the splice acceptor sequence, and wherein the open reading frame is in antisense orientation. 9. The method of claim 8, wherein the open reading frame encodes a marker gene. |
Details for Patent 7,205,148
Applicant | Tradename | Biologic Ingredient | Dosage Form | BLA | Approval Date | Patent No. | Expiredate |
---|---|---|---|---|---|---|---|
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | 06/04/1986 | ⤷ Try a Trial | 2039-02-26 |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | ⤷ Try a Trial | 2039-02-26 | |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | Injection | 103132 | ⤷ Try a Trial | 2039-02-26 | |
>Applicant | >Tradename | >Biologic Ingredient | >Dosage Form | >BLA | >Approval Date | >Patent No. | >Expiredate |
Make Better Decisions: Try a trial or see plans & pricing
Drugs may be covered by multiple patents or regulatory protections. All trademarks and applicant names are the property of their respective owners or licensors. Although great care is taken in the proper and correct provision of this service, thinkBiotech LLC does not accept any responsibility for possible consequences of errors or omissions in the provided data. The data presented herein is for information purposes only. There is no warranty that the data contained herein is error free. thinkBiotech performs no independent verification of facts as provided by public sources nor are attempts made to provide legal or investing advice. Any reliance on data provided herein is done solely at the discretion of the user. Users of this service are advised to seek professional advice and independent confirmation before considering acting on any of the provided information. thinkBiotech LLC reserves the right to amend, extend or withdraw any part or all of the offered service without notice.