WO1997033895A1 - Molecules d'arn isolees se fixant a l'arginine et leurs applications - Google Patents

Molecules d'arn isolees se fixant a l'arginine et leurs applications Download PDF

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Publication number
WO1997033895A1
WO1997033895A1 PCT/EP1997/001223 EP9701223W WO9733895A1 WO 1997033895 A1 WO1997033895 A1 WO 1997033895A1 EP 9701223 W EP9701223 W EP 9701223W WO 9733895 A1 WO9733895 A1 WO 9733895A1
Authority
WO
WIPO (PCT)
Prior art keywords
rna
amino acid
affinity
rna molecule
bound
Prior art date
Application number
PCT/EP1997/001223
Other languages
English (en)
Inventor
Albert Geiger
Michael Famulok
Original Assignee
Boehringer Mannheim Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Boehringer Mannheim Gmbh filed Critical Boehringer Mannheim Gmbh
Priority to JP9532275A priority Critical patent/JP2000507097A/ja
Priority to EP97915359A priority patent/EP0888367A1/fr
Publication of WO1997033895A1 publication Critical patent/WO1997033895A1/fr

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Classifications

    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07HSUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/30Preparation of optical isomers
    • C07C227/34Preparation of optical isomers by separation of optical isomers
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/38Separation; Purification; Stabilisation; Use of additives
    • C07C227/40Separation; Purification
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C273/00Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
    • C07C273/18Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups of substituted ureas
    • C07C273/189Purification, separation, stabilisation, use of additives
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07CACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C277/00Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
    • C07C277/08Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups of substituted guanidines
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C12N15/1006Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6811Selection methods for production or design of target specific oligonucleotides or binding molecules

Definitions

  • This invention relates to isolated RNA molecules with high or
  • the invention are processes lor securing these RNA molecules.
  • a third example is the interaction of the H ⁇ V-1 TAT protein with a stem loop structure of TAR RNA, located
  • TAR is a single arginine within a basic region of TAT.
  • RNA to largely determine specificity, functionality, and
  • RNA aptamers which specifically recognize amino acids
  • arginine specific aptamers might be especially relevant to protein-RNA
  • the HIV-1 Rev protein contains a basic region in which ten of seventeen amino acids between positions
  • RRE responsive element
  • TAR/ Tat complex an -irginine and an isoleucine residue were found to
  • Rex-protein of HTLV-I might interact with its natural
  • the present invention is a
  • Figure 1 is a representative elution protocol obtained in
  • the first pool (“pool 1" hereafter) was
  • pool 2 was then subjected to PCR amplification.
  • the PCR amplification was carrier out as described by Famulok, supra,
  • amplification product was extracted with phenol/ CHCL y precipitated with ethanol, dissolved in 1.3 ml of TE (lOmM Tris-HCl, pH 7.6; ImM
  • RNA produced in Example 1, supra was selected for high
  • the preselection column consisted of
  • RNA containing samples were added to the preselection column, and then washed with 2-3 volumes of buffer. The effect of this
  • the column was then eluted with another 5 column volumes of binding buffer containing 20mM citrulline so as to remove any non-
  • binding buffer plus 20mM arginine was then heat denatured, in
  • RNA typically
  • RNA molecules were used, i.e., Ag. 06 (SEQ ID NO: ), as well
  • the value for Ag. 06 was about 330nM, i.e., nearly 200 times greater
  • aminoglycoside antibiotics tobramycin, kanamycin, lividomycin
  • RNA ribonucleic acid
  • isolated ribonucleic acid molecules having
  • affinities of around 300nM, more preferably greater than about 60 ⁇ M are affinities of around 300nM, more preferably greater than about 60 ⁇ M.
  • RNA molecules can have the strong affinity for any L- or D-
  • RNA molecules are those set forth in SEQ ID NOS: 3 through 20 , inclusive.
  • RNA molecules have various uses, which will be clear to
  • RNA molecules may be used to
  • affinitv to form complexes of RNA and amino acid which can be
  • Enantiomers of amino acids may be identified and/or
  • amino acids such as L-arginine and L- ⁇ trulline may be separated, and
  • the bound RNA is contacted with a solution of an amino acid which is not the amino acid for which a high affinity binder is
  • steps using L-citrulline Preferably, one carries out the cycling for
  • affinity binder is sought. This serves to elute the desired RNA from
  • RNA be bound to the RNA
  • a strong or high affinity binder as was noted, supra, is an RNA molecule which has a low Kd value.
  • strong affinitv (or high affinity) binders are those with a Kd
  • these molecules have a Kd
  • this value is about 500 nM or less, and most

Abstract

L'invention, qui a trait à des molécules d'ARN isolées, dotées d'une affinité élevée pour des acides aminés particuliers, porte également sur des procédés permettant d'isoler ces molécules ainsi que sur leur utilisation. On isole ces molécules d'ARN par le biais d'une technique faisant intervenir différentes étapes de lavage et de réchauffage.
PCT/EP1997/001223 1996-03-12 1997-03-11 Molecules d'arn isolees se fixant a l'arginine et leurs applications WO1997033895A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP9532275A JP2000507097A (ja) 1996-03-12 1997-03-11 アルギニンに結合する単離されたrna分子およびその使用
EP97915359A EP0888367A1 (fr) 1996-03-12 1997-03-11 Molecules d'arn isolees se fixant a l'arginine et leurs applications

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US1411396P 1996-03-12 1996-03-12
US014,113 1996-03-12

Publications (1)

Publication Number Publication Date
WO1997033895A1 true WO1997033895A1 (fr) 1997-09-18

Family

ID=21763618

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1997/001223 WO1997033895A1 (fr) 1996-03-12 1997-03-11 Molecules d'arn isolees se fixant a l'arginine et leurs applications

Country Status (3)

Country Link
EP (1) EP0888367A1 (fr)
JP (1) JP2000507097A (fr)
WO (1) WO1997033895A1 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992014843A1 (fr) * 1991-02-21 1992-09-03 Gilead Sciences, Inc. Aptamere specifique de biomolecules et procede de production

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1992014843A1 (fr) * 1991-02-21 1992-09-03 Gilead Sciences, Inc. Aptamere specifique de biomolecules et procede de production

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
GEIGER A. ET AL.: "RNA Aptamers that bind L-arginine with sub-micromolar dissociation constants and high enantioselectivity" *
GEIGER, ALBERT ET AL: "RNA aptamers that bind L-arginine with sub-micromolar dissociation constants and high enantioselectivity", NUCLEIC ACIDS RESEARCH, vol. 24, no. 6, 15 March 1996 (1996-03-15), pages 1029 - 1036, XP002037703 *
GEIGER, ALBERT ET AL: "RNA aptamers that bind L-arginine with sub-micromolar dissociation constants and high enantioselectivity", NUCLEIC ACIDS RESEARCH, vol. 24, no. 6, 6 March 1996 (1996-03-06), XP002037702 *

Also Published As

Publication number Publication date
JP2000507097A (ja) 2000-06-13
EP0888367A1 (fr) 1999-01-07

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