The invention is directed to methods for synthesizing oligonucleotides direction on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3′-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3′ hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Provided herein are compounds that selectively bind and cleave RNA. In various embodiments, the disclosure provides chemical compounds effective as ribonuclease targeting chimeras (RIBOTACs), that target the endogenous enzyme RNase L to selectively cleave the RNA in a living cell. These compounds are useful in the treatment of diseases, e.g., the treatment of breast cancer.

The present disclosure relates to an aglycosylated antibody-producing transgenic mouse and the use of an aglycosylated antibody produced therefrom. When the transgenic mouse of the present disclosure is used, it is possible to easily produce aglycosylated antibodies against various target antigens, and to precisely diagnose disease by detecting a glycoprotein biomarker using the produced aglycosylated antibody.

The invention provides an antisense oligonucleotide reduced in toxicity. The antisense oligonucleotide has a central region, a 5-side region and a 3-side region, wherein the central region has a nucleotide (2-3 bridged nucleotide) in which the 2-position and the 3-position of a sugar moiety are bridged and/or a non-bridged nucleotide (3-position-modified non-bridged nucleotide) having a substituent at the 3-position.

Modification formats having modified nucleotides are provided for siRNA. Short interfering RNA having modification formats and modified nucleotides provided herein reduce off-target effects in RNA interference of endogenous genes. Further modification formatted siRNAs are demonstrated to be stabilized to nuclease-rich environments. Unexpectedly, increasing or maintaining strand bias, while necessary to maintain potency for endogenous RNA interference, is not sufficient for reducing off-target effects in cell biology assays.

The present disclosure provides oligomeric compound comprising a modified oligonucleotide having a central region comprising one or more modifications. In certain embodiments, the present disclosure provides oligomeric compounds having an improved therapeutic index or an increased maximum tolerated dose.

The invention is directed to methods for synthesizing oligonucleotides direction on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3 hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

The present application relates to nucleic acid compounds, compositions comprising same and methods of use thereof for treatment of various diseases, disorders and conditions. The compounds are preferably chemically synthesized and modified double-stranded nucleic acid molecules which down regulate expression of a p53 gene.

The present invention relates to RNAi molecules, and compositions thereof, comprising a 2 internucleoside linkage connecting the nucleotide at position 1 and the nucleotide at position 2 at the 5 end of the antisense strand. Specifically, the invention relates to single- and double-stranded short interfering nucleic acid (siNA) molecules that are capable of mediating RNA interference comprising 5 modified nucleotides that comprise, among other potential modifications, a 2 internucleoside linkage. The invention further relates to 5 modified nucleotides used as reagents to generate the RNAi molecules of the invention and methods of using the disclosed RNAi molecules.

Various embodiments provide STOPS polymers that are S-antigen transport inhibiting oligonucleotide polymers, processes for making them and methods of using them to treat diseases and conditions. In some embodiments the STOPS modified oligonucleotides include an at least partially phosphorothioated sequence of alternating A and C units having modifications as described herein. The sequence independent antiviral activity against hepatitis B of embodiments of STOPS modified oligonucleotides, as determined by HBsAg Secretion Assay, is greater than that of a reference compound.