Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 20 of 50 for “"guide RNA"”.
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GUIDE RNA-DEPENDENT AND INDEPENDENT tRNA MODIFICATIONS IN ARCHAEA
Stable RNAs undergo a wide variety of post-transcriptional modifications, that add to the functional repertoire of these molecules. Some of these modifications are catalyzed by stand-alone protein enzymes, while some others are catalyzed by RNA-protein complexes. tRNAs from all domains of life …
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Genetic circuits for functional screens of Cas12a guide RNA libraries
The discovery of CRISPR RNA-guided endonucleases have catalyzed huge technological advancements in the field of synthetic biology, such as the creation of gene drives: genomically encoded CRISPR systems capable of spreading through a wild population. These systems have two components: a …
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Cystic fibrosis gene repair: correction of ΔF508 using ZFN and CRISPR/Cas9 guide RNA gene editing tools
… nucleases designed in this lab and CRISPR/gRNAs designed in this thesis can mediate efficient homology-directed repair (HDR) with appropriate donor repair plasmids to correct CF-causing mutations in a CF cell line. Firstly, the most common mutation, ΔF508, was corrected using a pair of …
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STRUCTURAL AND FUNCTIONAL STUDIES OF ARCHAEAL BOX C/D GUIDE RNA AND ROLE OF A PUTATIVE HUMAN PSEUDOURIDINE SYNTHASE, PUS10 IN APOPTOSIS
RNAs undergo different posttranscriptional chemical modifications, which affect their structural stability and functional diversity. RNA methylation is a very common type of post-transcriptional modification and is present in all domains of life: Archaea, Eukaryotes and Bacteria. Some of these …
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Mechanism of Permissive Cleavage Activity of TevCas12a
… permissive cleavage. TevCas12a activity is guide RNA dependent, but not guide RNA sequence specific, it can degrade any DNA substrate with a non-targeted guide RNA. Sequencing of the TevCas12a cleavage products with Oxford Nanopore revealed that most of the non-guide directed cuts were …
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Genomic nucleic acid memory storage with directed endonucleases
… can recognize and cleave viral DNA using an RNA-guided directed endonuclease. Cuts left by the endonuclease are repaired by an error-prone DNA damage repair mechanism, namely non-homologous end joining (NHEJ), likely to leave mutations at the cut sites. Defining the cut site with …
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Precise and expansive genomic positioning for CRISPR edits
… tools has been fashioned from the natural DNA/RNA-targeting ability of certain CRISPR-associated (Cas) proteins and their guide RNA, which work together to recognize and defend against infectious genetic threats. This straight-forward RNA-programmed sequence recognition by CRISPR has …
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The Transposon-Encoded Protein TnpB is an RNA-Guided Nuclease
Programmable RNA-guided systems, such as CRISPR-Cas adaptive immune systems, perform a wide variety of biological functions and have been harnessed for important therapeutic and diagnostic biotechnologies. One CRISPR effector protein, the Cas12 nuclease, is hypothesized to have its evolutionary …
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The Interrogation of Cas9 Aptamers and sgRNA Structures Through SELEX
… of Cas9, the interrogation and evolution of sgRNA scaffold, in addition to novel Cas9 binding RNAs, represent another echelon of development and therapeutic potential. Currently, the majority of research utilizes a singular guide RNA scaffold sequence (the sgRNA) for a given Cas protein (e.g., …
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Focused CRISPR-Cas9 screens investigating the DNA damage response
… of three screens using focused CRISPR-Cas9 guide RNA libraries each interrogating different aspects of DDR biology. Firstly, we used a kinase-focused guide RNA library in combination with three clinically relevant DNA damaging agents. We identified kinase knockouts causing sensitivity and …
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Targeting nucleic acids for pancreatic cancer: disease modeling and therapy
… stromal barrier characteristic of PDAC. RNA interference through siRNA holds promise in targeting key mutations driving PDAC, such as oncogenic KRAS; however, a nucleic acid delivery vehicle that homes to PDAC and breaches the stroma does not yet exist. Noting that the novel cyclic …
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A TALE OF TWO METHYLATION MODIFICATIONS IN ARCHAEAL RNAs
<p>In all the three domains of life, most RNAs undergo post transcriptional modifications both on the bases as well as the ribose sugars of the individual ribonucleotides. 2'-O-methylation of ribose sugars and isomerization of Uridines to Pseudouridines are two most predominant modifications in …
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Discovery and biochemical characterization of RNA interference in budding yeast
RNA interference (RNAi) is a eukaryotic pathway for the post-transcriptional regulation of gene expression. In the simplest form of RNAi, a double-stranded RNA (dsRNA) trigger is converted into small-RNA duplexes by the Dicer enzyme. These duplexes are then loaded into the effector protein …
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Applications of genome editing for disease modeling in mice
… for understanding its function. The RNA-guided endonuclease Cas9 from microbial type II CRISPR (clustered regularly interspaced short palindromic repeat) systems has been harnessed to facilitate facile genetic manipulations in a variety of cell types and organisms. Cas9 can be easily …
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Machine learning based CRISPR gRNA design for therapeutic exon skipping
… We show that machine learning can select Cas9 guide RNAs that disrupt splice acceptors and cause the skipping of targeted exons. We experimentally measured the exon skipping frequencies of a diverse genome-integrated library of 791 splice sequences targeted by 1,063 guide RNAs in mouse …
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STRUCTURAL AND FUNCTIONAL CHARACTERIZATION OF ARCHAEAL BOX H/ACA RIBONUCLEOPROTEIN INVOLVED IN RIBOSOMAL RNA PSEUDOURIDYLATION
Ribosomal RNAs (rRNA) undergo several post-transcriptional modifications inside the cell. These modifications can be (1) RNA- independent (enzyme only) and (2) guide RNA-mediated. In the latter mechanism, a group of small, metabolically stable, non-coding RNAs, present as ribonucleoprotein (RNP) …
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Application of CRISPR/Cas9 to edit genes affecting seed morphology traits in wheat
… traits. CRISPR/Cas9 is based on a synthetic guide-RNA (gRNA) that can guide Cas9 nuclease to specific targets in the genome and create double strand breaks (DSB). The DSB are repaired through the error-prone non-homologous end joining process causing insertions or deletions that may result in …
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A novel CRISPR-Cas9 platform with divergent targeting capabilities
RNA-guided DNA endonucleases of the CRISPR-Cas system are widely used for genome engineering and thus have numerous applications in a wide variety of fields. The range of sequences that CRISPR endonucleases can recognize, however, is constrained by the need for a specific protospacer adjacent motif …
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An integrated CRISPR-Cas toolkit for engineering human cells
… from Streptococcus Pyogenes comprising of the RNA-guided DNA endonuclease Cas9 and the guide RNA (gRNA) has been demonstrated to enable unprecedented genome editing efficiency across all domains of life. Current applications however, employ CRISPR/Cas technology in a stand-alone fashion, …
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