Described herein are methods for the controlled de novo synthesis of RNA oligonucleotides using enzymatic catalysis. For example, provided herein are methods for preparing RNA oligonucleotides via controlled, template-independent addition of nucleotides to an initiator oligonucleotide 3′-terminus via enzymatic catalysis (also known as terminal transferase activity). Single nucleotides can be iteratively added by a compatible polymerase (e.g., a poly(N) polymerase such as a poly(U) polymerase) until a desired RNA oligonucleotide sequence is synthesized. Also provided are nucleotides and polymerases useful in the methods described herein.

The present invention relates to a pharmaceutical composition for preventing or treating cancer, comprising miR-324 and TUT4/7 expression modulators, wherein the pharmaceutical composition of the present invention can prevent cell division and hinder cancer development by inhibiting the function of TUT4/7, and can increase the amount of miR-324-5p and suppress the function of miR-324-3p.1, and therefore can be effectively used for prevention, treatment, or diagnosis of cancer.

This disclosure provides compositions comprising a modified oncolytic virus that can contain modifications in the viral genome and exogenous nucleic acids coding for proteins. The viral compositions and methods provided herein can be utilized for the treatment of cancer.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Described herein is a LIN28-independent role of TUTases in oncogenesis. Provided herein are compositions and methods for treating cancer via inhibition of TUTases. TUTase depletion also sensitizes the cells to disruptions in RNA metabolism and/or protein metabolism. Thus, further provided herein are strategies of combination therapy, combining TUTase inhibitors, agents that disrupt RNA metabolism, and agents that disrupt protein metabolism, to treat cancer.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between 3 end of the RNA and the polymer of non-canonical nucleotides to identify 30 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.