Vilnius University

Rwth Aachen

Biotechnologijos Institutas

Institute of Biotechnology

Saulius Klimasauskas

Edita Kriukiene

Zita Liutkeviciute

Zdislav Stasevskij

Elmar Weinhold

Christian Dalhoff

Grazvydas Lukinavicius

Giedrius Vilkaitis

Milda Mickute

Viktoras Masevicius

Rasa Rakauskaite

G Lukinavicius

Provided is a method for modifying a ssRNA at the 3' end, the method including contacting the strand with a ssRNA 2&#x2032;-O-methyltransferase in the presence of a co-factor, under conditions which allow for the transfer by the ssRNA 2&#x2032;-O-methyltransferase of a part of the co-factor onto the 3&#x2032; end of the ssRNA to form a modified ssRNA, wherein the ssRNA bears 2&#x2032;-OH group at 3&#x2032; terminal nucleotide and wherein the part of the co-factor transferred includes a reporter group or a functional group.

Analysis of single-stranded RNA

A method for producing a nucleic acid molecule from a template nucleic acid sequence and a linking unit attached to a primer, which method comprises a step of contacting the template nucleic acid sequence with a nucleic acid polymerase under conditions which allow the nucleic acid polymerase to produce the nucleic acid molecule from the primer based on the template nucleic acid sequence, wherein the linking unit is attached to a target site in the template nucleic acid sequence with a covalent linkage.

Nucleic acid production and sequence analysis

The present invention relates to targeted conversion of alpha-hydroxyalkylated residues in biomolecules in the presence of a directing methyltransferase, namely to targeted removal of the alpha-hydroxyalkyl moieties to give unmodified residues, or targeted derivatization of the alpha-hydroxyalkyl groups by covalent coupling of non-cofactor compounds represented by formula HQ-LX, wherein X represents a functional group or a reporter group attached via a linker moiety L, and QH is selected from HS&#x2014;, HSe&#x2014;, HO&#x2014;HN&#x2014;, HNor HCN in the presence of a directing methyltransferase. Further development of the method of targeted conversion comprises methods for targeted labeling a biomolecule and method for detecting hydroxymethylated target sites in a biomolecule according to the present invention.

Conversion of alpha-hydroxyalkylated residues in biomolecules using methyltransferases

The present invention relates to targeted conversion of alpha-hydroxyalkylated residues in biomolecules in the presence of a directing methyltransferase, namely to targeted removal of the alpha-hydroxyalkyl moieties to give unmodified residues, or targeted derivatization of the alpha-hydroxyalkyl groups by covalent coupling of non-cofactor compounds represented by formula HQ-LX1 wherein X represents a functional group or a reporter group attached via a linker moiety L, and QH is selected from HS&#x2014;, HSe&#x2014;, HO&#x2014;HN&#x2014;, HNor HCN in the presence of a directing methyltransferase. Further development of the method of targeted conversion comprises methods for targeted labeling a biomolecule and method for detecting hydroxymethylated target sites in a biomolecule according to the present invention.

The present invention relates to a use of non-cofactor compounds, represented by formulas (I) or (II) wherein R and Z are independently selected from H, D, C-C-alkyl, preferably C-C-alkyl, alkenyl, alkinyl, phenyl or -LX, wherein X represents a functional group or a reporter group attached via a linker group L, and QH is selected from &#x2014;SH, &#x2014;SeH, &#x2014;NHNHor &#x2014;ONH, for a targeted modification or derivatization of a biomolecule by covalent coupling to the biomolecule in the presence of a directing methyltransferase. Further development of the method of targeted modification and derivatization are the method for targeted labeling a biomolecule and method for detecting unmethylated target sites in a biomolecule comprising modification of the biomolecule according to the present invention. 

Derivatization of biomolecules by covalent coupling of non-cofactor compounds using methyltransferases

S-Adenosyl-L-methionine analogs of formula (I): are disclosed wherein R comprises a carbon-carbon double bond, carbon-sulfur double bond, carbon-nitrogen double bond, -a carbon-carbon triple bond, carbon-nitrogen triple bond or an aromatic carbocyclic or heterocyclic system in &#x3b2;-position to the sulfonium center, Xis an organic or inorganic anion carrying one or more negative charges, Z is &#x2014;CRR&#x2014;, &#x2014;O&#x2014;, &#x2014;S&#x2014; or &#x2014;NR&#x2014; and R, Rand Rare independently selected from H, D and C&#x2014;C alkyl; as well as complexes with a methyltransferase, pharmaceutical compositions, methods for modifying a biomolecule, and methods for detecting sequences specific methylation of biomolecules.

Growing community of inventors

Vilnius, Lithuania

Saulius Klimasauskas