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Engineered RNA viral synthesis of microRNAs - …

Several strategies have been employed to mitigate CRISPR’s propensity for off-target genome modification. One such strategy is to use double nickases to create DSBs. The Cas9 D10A mutant is able to cleave only one DNA strand, thereby creating a “nick”. When two sgRNAs that bind on opposite strands flanking the target are introduced, two Cas9 D10A nickase miolecules together create a staggered-cut DSB, which is then repaired by either NHEJ or HR (Figure 3). The double nickase strategy has been shown to greatly reduce the frequency of off-target modification. However, double nickases are limited in utility by design constraints; the sgRNAs must be on opposite strands, in opposite orientation to one another, and display optimal activity when spaced from 3-20 nucleotides apart. In addition, the cleavage activity of double nickases tends to be lower than that of standard Cas9:sgRNA. Further, nickases can still cause some degree of off-target indel formation.

Engineered RNA viral synthesis of microRNAs ..

Shu-Bing Qian researches mRNA translation and the mechanisms that regulate the onset of protein synthesis under certain cellular conditions. A clearer conception of the factors that control the quality and quantity of protein production during events such as cell growth, differentiation, or stress response could help define new therapeutic strategies for diseases such as cancer, diabetes, and neurodegenerative diseases.

Engineered RNA viral synthesis of microRNAs - Europe …

Engineered RNA viral synthesis of microRNAs Andrew Varblea,b,1, Mark A

Francesca Storici is studying how cells use RNA as a template to repair DNA lesions in a process in which genetic information flows in reverse from RNA to DNA. Furthermore, during DNA metabolism, subunits of RNA can be incorporated into the genome, and Storici is investigating spectra, consequences, and whether these RNA intrusions are associated with cellular stress and/or cancer.

Francesca Storici is studying how cells use RNA as a template to repair DNA lesions in a process in which genetic information flows in reverse from RNA to DNA. Furthermore, during DNA metabolism, subunits of RNA can be incorporated into the genome, and Storici is investigating spectra, consequences, and whether these RNA intrusions are associated with cellular stress and/or cancer.

Engineered RNA viral synthesis of microRNAs - CORE

Engineered RNA viral synthesis of microRNAs: Authors: ..

Award amounts will be determined by HHMI, the Bill & Melinda Gates Foundation, and the Simons Foundation based on several factors including award recipients’ level of external funding at the time of the award. Each award recipient’s institution will be given an additional 20% of the yearly grant for indirect (or administrative) costs.

In the type II CRISPR systems, the complex of a CRISPR RNA (crRNA) annealed to a trans-activating crRNA (tracrRNA) guides the Cas9 endonuclease to a specific genomic sequence, thereby generating double-strand breaks (DSBs) in target DNA. This system has been simplified by fusing crRNA and tracrRNA sequences to produce a synthetic, chimeric single-guided RNA (sgRNA). The sgRNA contains within it a 20 nucleotinde DNA recognition sequence (Figure 1).

(2010) Engineered RNA viral synthesis of microRNAs
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Engineered RNA viral synthesis of microRNAs.

Oregon State University, Corvallis, OR
Research Assistant, Dr. J.D. White | 1984 – 1987
Pharmaceutical synthesis; structure determination of protein and natural products; molecular modeling; drug delivery, particularly prodrug synthesis.

Noncanonical cytoplasmic processing of viral microRNAs RNA 2010 16 ..

Mr. Scotti’s practice includes all facets of intellectual property practice including patents, opinions, clearance, and litigation. He has worked in both the U.S. and Europe on patent drafting, patent litigation, and trademark prosecution. His extensive experience in both the U.S. and Europe gives him a unique perspective on the international protection of intellectual property. Mr. Scotti has lectured in both the U.S. and Europe on how to draft patent applications for global protection. He has also assisted in European litigations with issues that intermingle U.S. and European law.

PLOS Pathogens: Virus-Encoded microRNAs: An Overview and …

Shu-Bing Qian researches mRNA translation and the mechanisms that regulate the onset of protein synthesis under certain cellular conditions. A clearer conception of the factors that control the quality and quantity of protein production during events such as cell growth, differentiation, or stress response could help define new therapeutic strategies for diseases such as cancer, diabetes, and neurodegenerative diseases.

Proc Natl Acad Sci USA 107: 11519–11524.

Answer: The presence of the CMV or other promoters driving expression of the Cas9 coding sequences permits expression from the plasmid DNA. sgRNA transcription is driven by the U6 promoter. We recommend that you use the most efficient method for your cell type of interest. The following are all acceptable approaches for delivering our genome editing tools into cells:

Rabies Virus Infection and MicroRNAs - ScienceDirect

Biomedical researchers are enjoying a Renaissance in functional genomics, which aims to use a wealth of DNA sequence information—most notably, the complete sequence of the human genome—to determine the natural roles of the genes encoded by the genome. As a result, biochemical networks and pathways will be better understood, with the hope of leading to improved disease treatments. Researchers are turning increasingly to CRISPR (clustered, regularly interspaced, short palindromic repeats) for functional genomics studies. Several groups recently adapted CRISPR for high-throughput knockout applications, by developing large-scale CRISPR sgRNA libraries. GeneCopoeia recently launched a number of smaller, pathway- and gene group-focused CRISPR sgRNA libraries, which offer several key advantages over the whole-genome libraries. In this 40 minute webinar, we discuss the merits and applications for CRISPR sgRNA libraries, how to use CRISPR sgRNA libraries, the advantages of using small, pathway- and gene group-focused libraries, and how GeneCopoeia can help you with your high-throughput CRISPR knockout studies.

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