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 18 of 18 for “"crRNA"”.
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Methods for CRISPR Cas12a Multiplexing in Mammalian Systems
… nucleic acid sequence through the binding of a crRNA hairpin. In recent years, these proteins have been adapted for a number of gene editing applications. The fusion of additional proteins of interest to dead nuclease (non-cutting) mutants of CRISPR proteins such as Cas9 and Cas12a have allowed …
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DESIGN AND INVESTIGATION OF DUMBBELL-SHAPED DNA MINIMAL VECTORS FOR CRISPR/CAS9 EDITING
… structure revealed that unstructured CRISPR RNA (crRNA) domains in sgRNA exhibited higher target cleavage activities as compared with structured crRNA domains. We also found that molecular helper functions enhancing DNA nuclear import (SV40 enhancer sequence) and Cas9 mRNA nuclear export (WPRE), …
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PART I CRYSTAL STRUCTURE OF A DIMERIZATION DOMAIN OF DROSOPHILA CAPRIN. PART II CHARACTERIZATION OF TWO CAS13B CRISPR-CAS SYSTEMS FROM PORPHYROMONAS GINGIVALIS
… EFFECTOR PROTEINS ARE CAPABLE OF PROCESSING PRE-CRRNA INTO MATURE CRRNA CRISPR-Cas adaptive immune system protects microorganism from foreign nucleic acids invasion through endonucleases activity guided by RNA, which system has turned to a powerful genome editing tool applied to a multifold …
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Desarrollo de una herramienta software de identificación de secuencias patógenas candidatas para el diseño de RNAs guía en el sistema SHERLOCK de diagnóstico
… un patógeno, a través de las cuales se diseñarán crRNAs, para ser utilizados en la detección de dicho organismo mediante la construcción de un test SHERLOCK con este crRNA.
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Investigation of Diverse CARF Effectors in Type III CRISPR - Cas Immunity
… array, generating guide RNA sequences called crRNA. These crRNA will associate with a CRISPR nuclease to create an RNA-guided nuclease that mediates site-specific degradation of invader DNA or RNA that is complementary to the crRNA. The type III CRISPR-Cas system is also able to synthesize a …
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Discovery and biochemical characterization of Casl3b, a Type VI-B CRISPR-Associated RNA-Guided RNase
… Casl3b processes its own CRISPR array(s) into crRNA(s) and Casl3b with either crRNA architecture targets single stranded RNA cleavage using its HEPN domains. SsRNA cleavage occurs at pyrimidine residues and is constrained by 5' and 3' protospacer-flanking sequences on the target. Casl3b also …
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CRISPR-integrated Biosensors for Human Health: Novel strategies for pathogen detection and cancer screening
… covalent crosslinking between Cas12a and crRNA, which improved detection of SARS-CoV-2 in clinical samples. Together, these studies demonstrate the potential of CRISPR-based biosensors as powerful tools for diagnostics. The systems developed in this work are adaptable, sensitive, and …
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Engineering programmable nucleases for in vivo and in vitro applications
… incorporates 2-aminoadenine (base Z) into the crRNA. This modification enhances the binding affinity between the crRNA and its complementary DNA target, resulting in a highly efficient and precise genome editing system. zCRISPR-Cas12a exhibits on-target editing efficiency comparable to that of …
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Discovery and characterization of Cas13b, a differentially regulated RNA-targeting CRISPR system
… effector protein and programmable CRISPR RNA (crRNA) to target nucleic acids, represent the most promising tool to target RNA. Building on previous research, a biocomputational pipeline was developed to discover novel functional class 2 CRISPR systems lacking the canonical adaptive machinery of …
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Bacterial defense islands: CRISPR-Cas and toxin-antitoxin systems /
… complexes can assemble on different length of crRNA and enable dsDNA targeting in a WT-like manner. The second part of the thesis is dedicated to the biochemical and structural characterization of the toxin-antitoxin system HEPN-MNT that is present in the operon of the type I-D CRISPR-Cas …
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Characterization and Engineering of Transposons for Genome Editing
… type V-K systems use a non-canonical CRISPR RNA (crRNA) to perform this task. Surprisingly, the distinct type I-B CAST uses a dedicated sequence-specific DNA binding protein for homing to an attachment site. Next, we engineer a non-long terminal repeat (LTR) retrotransposon, R2 from the silkworm …
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FtsK Regulates the Assembly of the Division Machinery in the FtsZ-less Chlamydia trachomatis
… cell division. In addition, CRISPRi which uses a crRNA to target an inducible and defective Cas12 enzyme (dCas12) to a specific chromosomal sequence, has shown that chlamydial FtsK also colocalizes with a chromosomal sequence immediately adjacent to the dif site during cell division. However, when …
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Discovery of novel systems for genome editing
… assay, we show that CASTs use CRISPR RNAs (crRNAs) from these defense systems for horizontal gene transfer. A high-resolution structure of the type I-F CAST-Cascade in complex with a type III-B crRNA reveals that Cas6 recognizes direct repeats via sequence-independent π − π interactions. In …
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Desenvolvimento de protocolo de edição gênica de Enterococos resistentes à vancomicina (VRE) empregando CRISPR-Cas
… um melhor resultado. Foi utilizado um crRNA específico previamente desenhado para o gene de resistência. Para a inativação do gene alvo, foi utilizado o sistema Alt-R® CRISPR-Cas9 e para formação do complexo RNP foi utilizado o sistema Alt-R® CRISPR-Cas9 RNP. A entrega do sistema de …
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Uncovering New Mechanisms of Type III CRISPR-Cas System Inhibition and Effector Function
… complex that functions together with its crRNA in target binding and cleavage. During type III CRISPR-Cas immunity in prokaryotes, RNA-guided recognition of viral (phage) transcripts stimulates the Cas10 complex to convert ATP into cyclic oligoadenylates. These act as signaling molecules …
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CPISPR-CAS: From a Prokaryotic Immune System to a Gene Editing Tool
… specificity is programmed by small CRSPR RNAs (crRNAs). In collaboration with Dr. David Bikard, a former colleague, we found when reprogrammed to target the genomes of host bacteria, CRISPR can target and kill the cells. The lethal consequence upon targeting made CRISPR-Cas a novel tool for …
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Identification and Characterization of Novel CRISPR-Cas Immune Effectors
… and maturation of CRISPR arrays into crRNAs containing spacer sequences that are subsequently loaded onto a CRISPR effector protein or protein complex that survey nucleic acids for sequences complementary to transcribed spacer sequences (protospacers). Upon base pairing between crRNAs …