One step engineering of the small-subunit ribosomal RNA using CRISPR/Cas9
10 Pages · 2016 · 752 KB · English
quintessential for understanding gene functions. Here we demonstrate the engineering of the small- ribosomal subunit (16S) RNA of Mycoplasma mycoides, by combining the CRISPR/Cas9 system and the yeast recombination machinery. We cloned the entire genome of M. mycoides in yeast and used.
Axonal target specificity in the CRISPR/Cas9 era
170 Pages · 2017 · 31.29 MB · English
Axonal target specificity in the CRISPR/Cas9 era : a new role for Reelin in vertebrate visual sytem young guy (the lab) and the real one (me) grow scientifically and personally. I am grateful in the last part of the introduction I will describe the CRISPR/ Cas9 system and the technologies develop
CRISPR-Cas9 Mediated Telomere Removal Leads to Mitochondrial Stress and Protein Aggregation
14 Pages · 2017 · 1.7 MB · English
Supporting the pathological relevance of cell aging by CRISPR-Cas9 mediated telomere removal, alterations were .. DNA loci were obtained from SYBR Green based real-time PCR using QuantStudio 6 flex Real-Time. PCR System . using the Pierce™ BCA protein assay kit (ThermoScientific).
One-step generation of triple gene-targeted pigs using CRISPR/Cas9 system
7 Pages · 2016 · 1.09 MB · English
Received: 02 October 2015. Accepted: 08 Thus, it drives us to recapitulate some key features of PD disease by generating DJ-1/parkin/PINK1 triple-gene modified pigs using. CRISPR/Cas9 system. Results .. analyzed by confocal microscopy using a LSM 5 PASCAL (ZEISS, Oberkochen, Germany).
Genome engineering using the CRISPR-Cas9 system
28 Pages · 2013 · 1.67 MB · English
cleavage, we further describe a double-nicking strategy using the cas9 nickase mutant with paired guide rnas. this protocol provides experimentally derived guidelines for the selection of target sites, evaluation of cleavage efficiency and analysis of off-target activity. Beginning with target desi
Exciting Prospects for Precise Engineering of Caenorhabditis elegans Genomes with CRISPR/Cas9
8 Pages · 2013 · 672 KB · English
Biomedical Sciences and Danish National Research Foundation Centre for Cardiac Arrhythmia, University of Copenhagen,. 2200 Kbh N . by oligonucleotides and was shown to function in further diverged nematode .. nematodes other than C. elegans because a detailed knowl- edge of active
Characterizing a thermostable Cas9 for bacterial genome editing and silencing
11 Pages · 2017 · 1.81 MB · English
Mihris I.S. Naduthodi1, Alex Gussak1, Rudolf B.L. Brinkman2, Richard van Kranenburg 1,2 & John van der Oost1. CRISPR-Cas9-based genome engineering tools have selected for culturing under microaerobic lactate-producing conditions for 24h44. The growth of the pThermoCas9i_ldhL cultures
Small Molecule-Triggered Cas9 Protein with Improved Genome
10 Pages · 2015 · 741 KB · English
thereby enhancing genome-modification specificity. Together with our recent observations that direct delivery of transient Cas9 protein into cells can also improve its specificity16, these findings strongly suggest that limiting the opportunity of genome-editing agents to modify off-target loci fol
Efficient Gene Knock-out and Knock-in with Transgenic Cas9 in Drosophila
5 Pages · 2014 · 869 KB · English
developing a transgenic system that expresses Cas9 in the Drosophila germline. Using this system, we induced inheritable knock-out mutations by injecting only the gRNA into embryos, achieved highly efficient mutagenesis by expressing gRNA from the promoter of a novel non-coding RNA gene, and
CRISPR/Cas9-Mediated Genome Editing in Soybean Hairy Roots
13 Pages · 2015 · 2.08 MB · English
CRISPR/Cas9 system to simultaneously edit two endogenous soybean genes using only one customized genome modifications in target genes of soybean hairy roots could rapidly assess the effi- ciency of .. firmed the presence of indel mutations at the two target sites (S2C and S3C Figs). Multiple
Could the CRISPR/Cas9 System be applied to Atherosclerosis?
4 Pages · 2016 · 1.08 MB · English
to inactivate harmful genes. Additionally, once CRISPR/Cas9 is proposed for the embryonic stage in animal models, the progeny might generate.
CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes
10 Pages · 2015 · 1.19 MB · English
technologies such as CRISPR/Cas9 in human pre-im- plantation embryos. In this report, we used tripronuclear. (3PN) zygotes to further investigate CRISPR/Cas9-me- diated gene editing in human cells. We found that. CRISPR/Cas9 could effectively cleave the endogenous β-globin gene (HBB).
How specific is CRISPR/Cas9 really? Henriette O'Geen, Abigail S. Yu and David J. Segal* Genome ...
20 Pages · 2016 · 1.21 MB · English
Abstract. The specificity of RNA-guided nucleases has gathered considerable interest as they become broadly applied to basic research and therapeutic development. Reports of the simple generation of animal models and genome engineering of cells raised questions about targeting precision.
Combining CRISPR/Cas9 mutagenesis and a small-molecule inhibitor to probe the function of ...
36 Pages · 2017 · 2.83 MB · English
cancer cell lines harboring CRISPR/Cas9-induced null mutations in MELK exhibit wild-type growth in vitro, under environmental experimental interpretation (23, 24), CRISPR/Cas9 mutagenesis is also prone to several important limitations. Stem Cells Dayt Ohio 31(5):870–881. 22. Hebbard L, et al.