144results about How to "Reduce off-target effects" patented technology

The invention relates to a double-stranded RNA (dsRNA) molecule comprising between about 15 base pairs and about 40 base pairs, at least one 5′-methyl-pyrimidine and at least one 2′—O-methyl ribonucleotide, and a methods of using such modified dsRNA molecule to increase stability of a siRNA molecule when in contact with a biological sample, to reduce off-target effects and to reduce interferon responsiveness (IFN) of the siRNA molecule.

The invention aims at the technical defects of off-target effects of an existing CRISPR / Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR associated protein 9) gene editing technology to carry out site-directed mutation of an amino acid site on a site combined with a DNA (Deoxyribonucleic Acid) sequence of Cas9 protein to obtain the Cas9 protein with low off-target efficiency,so as to realize a gene editing unction with higher targeting performance and higher efficiency. Meanwhile, the invention further provides a method for carrying out HEK293T cell gene editing by adopting a gene editing method.

The present invention generally provides vectors, compositions, and methods of using the same for gene therapy.

A hemi-sulfate salt of the structure:to treat a host infected with hepatitis C, as well as pharmaceutical compositions and dosage forms, including solid dosage forms, thereof.

The invention belongs to the field of nucleic acid editing, and particularly relates to the technical field of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR). In particular, the present invention provides Cas effector proteins, fusion proteins comprising such proteins, and nucleic acid molecules encoding them, and also provides complexes and compositions for nucleic acid editing (e.g., gene or genome editing) comprising the aforementioned proteins or nucleic acid molecules, and methods for nucleic acid editing (e.g., gene or genome editing) comprising the aforementioned proteins.

This invention encompasses compounds, structures, compositions and methods for therapeutic guide molecules that direct CRISPR gene editing. A guide molecule for directing gene editing can be allele selective, or disease allele selective, and can exhibit reduced off target activity. A guide molecule can be composed of monomers, including UNA monomers, nucleic acid monomers, and modified nucleotides, wherein the compound is targeted to a genomic DNA. The guide molecules of this invention can be used as active ingredients for editing or disrupting a gene in vitro, ex vivo, or in vivo.

The invention discloses an oligonucleotide molecule used for inhibiting expression of mRNA of a target gene and a composition set thereof, and provides siRNA which is composed a positive-sense strand which is composed of 19-27 nucleotides and a negative-sense strand being reverse-complementary with the positive-sense strand. In the positive-sense strand, both 5-9 continuous nucleotides from a 5'-terminal and 5-9 continuous nucleotides from a 3'-terminal are 2'-ribose modified nucleotides. The siRNA molecular mixture can influence expression of target genes in at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% and 90% cells, and is at least 45%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% and 95% in inhibition ratio.

The invention relates to biological technology, specifically relates to a chimeric protein pAgoE, a construction method and applications thereof, and at the same time, relates to a chimeric protein pAgoE using a guide, a construction method and applications thereof, and a chimeric protein, which is based on the chimeric protein pAgoE and has a genome targeting editing function. The chimeric protein pAgoE contains a pAgo structural domain with a genome targeting positioning function and an effect structural domain E, which is connected to the pAgo structural domain and has a genome cutting or modifying activity. The pAgo structural domain with a genome targeting positioning function and the effect structural domain E with a genome cutting or modifying activity are fused to establish the chimeric protein pAgoE; the chimeric protein pAgoE maintains the original targeting binding performance of the genome (DNA) having a targeting positioning function in the pAgo structural domain and the genome catalytic or modifying performance of the effect structural domain E at the same time, and an sufficient space is provided for the structural domain E to fully exert the effect.

The invention discloses an MCIR (MelanoCortin 1 Receptor) gene carrier and a construction method thereof. The MCIR gene carrier comprises MC1R-neo and MC1R-GFP (Green Fluorescent Protein), wherein the nucleotide sequence of the MC1R-neo is SEQ ID NO:1, and the nucleotide sequence of the MC1R-GFP is SEQ ID NO:2. A CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 system is adopted to construct an MC1R gene knock-out vector, and only a sgRNA (Ribonucleic Acid) of which the length is about 20bp needs to be designed by aiming at a gene knock-out point to be connected with a universal Cas9 gene. Therefore, compared with traditional gene knock-out technologies including ZFNs (Zinc finger nuclease), TALENs (Transcription Activator-Like Effector Nucleases) and the like, the construction method disclosed by the invention adopting the CRISPR / Cas9 to construct the gene knock-out vector is simpler and more convenient and is more convenient to be further popularized and applied to subsequent experiments.

This invention provides UNA oligomers for gene silencing with reduced off-target effects. The UNA oligomers can have a first strand and a second strand, each of the strands being 19-29 monomers in length, the monomers being UNA monomers and various nucleic acid monomers. Embodiments include pharmaceutical compositions and methods for treating or preventing TTR-related amyloidosis with reduced off-target effects by administering a UNA oligomer to a subject.

The invention provides a chemically modified double-strand small interfering RNA (siRNA) molecule and its preparation and application methods. Chemically modified nucleotide is introduced into the 14th site or / and 16th site beginning from 5' terminal of the siRNA molecular sense strand, thereby reducing the off-target effect caused by the modified siRNA molecular sense strand and enhancing specificity of the modified siRNA molecule for silence of target gene expression.

The present disclosure is directed to nanoconjugates that cross the blood-brain barrier and methods of their therapeutic use.

The invention provides a cell strain capable of reducing replicability adenovirus production and a construction method and an application thereof. The cell strain HEK 293.CS capable of reducing replicability adenovirus production is a safe adenovirus production cell line by knocking out of a gene fragment in HEK293 which is homologous with an Ad5 adenovirus E1 gene, and simultaneously providing template plasmid for replacing the gene fragment to a non-homologous sequence which is capable of stabilizing the E1 gene expression. Compared with un-reconstructed HEK293 cell strain, the growth capability and virus production capability of HEK 293.CS are not decreased, but detectable RCA is not produced; the cell strain can be used for large-scale cultivation of recombinant human adenovirus type 5, and the RCA production probability during medicine manufacture such as vaccines and antibodies is reduced.

The present invention provides immunoresponsive cells, including T cells, cytotoxic T cells, regulatory T cells, and Natural Killer (NK) cells, expressing an antigen recognizing receptor and an inhibitory chimeric antigen receptor (iCAR). Methods of using the immunoresponsive cell include those for the treatment of neoplasia and other pathologies where an increase in an antigen-specific immune response is desired.