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.

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 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.