Compositions and methods for cell transformation

• US 5,795,870 A
• Filed: 11/07/1994
• Issued: 08/18/1998
• Est. Priority Date: 12/13/1991
• Status: Expired due to Fees

• Current Assignee: The Trustees of Princeton University (Princeton University)
• Original Assignee: The Trustees of Princeton University (Princeton University)
• Inventors: Kahne, Suzanne Walker
• Primary Examiner(s): Kight, John
• Assistant Examiner(s): Lee, Howard C.
• Application Number:
  US08/336,675
• Time in Patent Office: 1,380 Days
• Field of Search: 514/26, 536/5
• US Class Current: 514/26
• CPC Class Codes: A61K 31/575 substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: C07J 17/005 Glycosides
• CPC Class Codes: C07J 31/006 not covered by C07J31/003
• CPC Class Codes: C07J 41/0005 the nitrogen atom being directly linked to the cyclopenta(a)hydro phenanthrene skeleton
• CPC Class Codes: C07J 41/0011 Unsubstituted amino radicals
• CPC Class Codes: C07J 41/0027 Azides
• CPC Class Codes: C07J 41/0061 one of the carbon atoms being part of an amide group
• CPC Class Codes: C07J 43/003 not condensed
• CPC Class Codes: C07J 51/00 Normal steroids with unmodified cyclopenta(a)hydrophenanthrene skeleton not provided for in groups C07J1/00 - C07J43/00
• CPC Class Codes: C07J 9/005 containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol

Abstract

The present invention relates to methods and compositions for the transformation of cells. In particular, compositions and methods are disclosed which include combinations of the nucleic acid of interest and polyhydroxylated or polyglycosylated steroid molecules. Most preferably, exogenous or endogenous nucleic acid is contacted with the cell in the presence of a bile acid (e.g., cholic acid) derivatized with an amine-containing side chain.

Claims

1. A method of enhancing the transformability of a host cell comprising contacting a host cell with a compound of the formula (I):

##STR2## in which R₁ can be an H, OH, OR₅, NH₂, NHR₆ or NR₆ R₇ ;

R₂ and R₃ may be the same or different and can be an H, OH or OR₅ ;

R₄ can be CONH₂, CONHR₆, CONR₆ R₇, CH₂ NH₂, CH₂ NHR₆, CH₂ NR₆ R₇, CO₂ --Y--NH₂, CO₂ --Y--NHR₆, or CO₂ --Y--NR₆ R₇ ;

R₅ is a protected or unprotected glycosyl moiety comprising 1-10 monosaccharide units in which the glycosidic linkage at the anomeric carbon atom of each monosaccharide unit is independently alpha or beta;

NH₂, NHR₆, and NR₆ R₇ represent an unsubstituted amino group, a monosubstituted amino group, and a disubstituted amino group, respectively, in which R₆ and R₇ may be the same or different and represent a hydrocarbon group comprising 1-15 carbon atoms substituted with one or more unsubstituted, monosubstituted or disubstituted amino groups;

Y represents a linear or branched alkylene group comprising 1-10 carbon atoms;

n is an integer from 0-10;

or its acid addition or quaternary ammonium salt;

provided that said compound comprises at least one R₆ group as defined above; and

(b) allowing said compound to remain in contact with said host cell while exposing said host cell to transforming nucleic acid.

2. The method of claim 1 in which said hydrocarbon group comprises a linear or branched aliphatic group.

3. The method of claim 1 in which said hydrocarbon group is cyclic.