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Potential high-frequency off-target mutagenesis induced by CRISPR/Cas9 in <Emphasis Type=

CRISPR/Cas9 tools has been a major concern along with the reports of their successful applications. We report unexpected observations of high frequency off-target mutagenesis induced by CRISPR/Cas9 in T1 ... applications (Wolt et al. 2016) . High-frequency off-target mutagenesis induced by CRISPR/Cas9 has been reported in human cells (Fu et al. 2013, Hsu et al. 2013; O’Geen et al. 2015; Tsai and Joung 2016

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potential therapeutic target value. H19; Prognosis; Surveillance; ID2; AML Background Acute myeloid leukemia (AML), the most common adult leukemia, is an etiologically, clinically, cytogenetically, and ... significance in AML remain poorly determined. Herein, we reported H19 as a prognostic and predictive biomarker in AML, and H19 played a crucial role in leukemogenesis with potential therapeutic target value

Chromatin accessibility is associated with CRISPR-Cas9 efficiency in the zebrafish (Danio rerio)

CRISPR-Cas9 technology is routinely applied for targeted mutagenesis in model organisms and cell lines. Recent studies indicate that the prokaryotic CRISPR-Cas9 system is affected by eukaryotic ... vivo activity. This suggests that factors present in vivo prevent Cas9 from acting on its target site. Fig 1. In vitro and in vivo CRISPR-Cas9 mutagenesis efficiencies do not correlate for all genes. a

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

type II system adapted from Streptococcus pyogenes has been the most exploited as a tool for genome engineering and gene regulation. In this review, we describe the different applications of CRISPR/Cas9 ... 532 ( 7600 ): 517 - 521 27. Fu Y , Foden JA , Khayter C , Maeder ML , Reyon D , Joung JK , Sander JD ( 2013 ) High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells . Nat

An optimized electroporation approach for efficient CRISPR/Cas9 genome editing in murine zygotes

Electroporation of zygotes represents a rapid alternative to the elaborate pronuclear injection procedure for CRISPR/Cas9-mediated genome editing in mice. However, current protocols for ... TransferMan NK2 micromanipulators (Eppendorf). The CRISPR/Cas9 solution was injected into the male pronucleus using injection capillaries (BioMedical Instruments, BM100F-10; type PI-1.6). If not stated

Targeted genome engineering in human induced pluripotent stem cells from patients with hemophilia B using the CRISPR-Cas9 system

. Highfrequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells . Nat Biotechnol . 2013 ; 31 ( 9 ): 822 - 6 . 45. Pattanayak V , Lin S , Guilinger JP , Ma E , Doudna JA , Liu DR . High ... . Off-target detection of CRISPR-Cas9 Potential off-target sites that differed from the sgRNA sequence by up to five nucleotides in the genome were searched using the web-based program Cas-OFFinder (http

The CRISPR/Cas9 system sheds new lights on the biology of protozoan parasites

The CRISPR/Cas9 system, a natural defence system of bacterial organisms, has recently been used to modify genomes of the most important protozoa parasites. Successful genome manipulations with the ... activity of the natural CRISPR/Cas9 type II system differs from that used in the genome engineering. To change the genome, it is necessary to provide the Cas9 enzyme and sgRNA (single-guide RNA) scaffold in

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were introduced. We recorded a high proportion of untouched cells in controls (median, 80%) compared to manipulated cells in which mites had been introduced (median, 12.5%) with a significant difference ... brood cells, hygienic bees were more responsive in the high mite density regime of eight and ten mites, an indication of a possibility that chemicals play a role in identification of Varroa -infested

Efficient CRISPR/Cas9-based genome editing in carrot cells

health-promoting crop grown worldwide. In this study, for the first time, we report application of the CRISPR/Cas9 system for efficient targeted mutagenesis of the carrot genome. Multiplexing CRISPR/Cas9 ... efficiently for genome engineering in Arabidopsis thaliana . Plant J 79 ( 2 ): 348 - 359 Fu Y , Foden JA , Khayter C , Maeder ML , Reyon D , Joung JK , Sander D ( 2013 ) High-frequency off-target mutagenesis

CRISPR/Cas9-mediated targeted mutagenesis in grape

for wine. Here, we report successful targeted mutagenesis in grape (Vitis vinifera L., cv. Neo Muscat) using the CRISPR/Cas9 system. When a Cas9 expression construct was transformed to embryonic calli ... specific target locus in independent cells, meaning that the limitations of low transformation efficiency can be overcome by the high performance of SSNs. Of the SSNs now available, the CRISPR/Cas9 system

Effects of <Emphasis Type=

The naturally occurring Verticillium nonalfalfae shows promise for biocontrol of the highly invasive Tree of Heaven (Ailanthus altissima), but might also bear a risk for non-target tree species. In ... indigenous non-target tree species, often associated with Tree of Heaven, and two invasive exotic tree species. In our study, V. nonalfalfae induced characteristic wilting symptoms and high mortality only on

Development of a multiple-gene-loading method by combining multi-integration system-equipped mouse artificial chromosome vector and CRISPR-Cas9

available for the selection of multiple-GLV integration. To overcome this issue, we attempted to knock out and reuse drug resistance genes (DRGs) using the CRISPR-Cas9 system. In this study, we developed new ... -target issues with the CRISPR-Cas9 system: Since genome editing might cause unexpected mutations, more effort is required to eliminate the possibility of off-target effects on host genomes. ( 5

Effects of <Emphasis Type=

Effects of Rhizophora mangle leaf litter and seedlings on carbon and nitrogen cycling in salt marshes - potential consequences of climate-induced mangrove migration Hendrikus J. Laanbroek 0 1 2 3 ... conditions, potential nitrification and denitrification activities and abundances of genes that are related to microbial processes in the nitrogen cycle. Results Growth of R. mangle seedlings was stimulated in

Differences between immunodeficient mice generated by classical gene targeting and CRISPR/Cas9-mediated gene knockout

Immunodeficient mice are widely used for pre-clinical studies to understand various human diseases. Here, we report the generation of four immunodeficient mouse models using CRISPR/Cas9 system ... target gene that may lead to frameshift mutations (Lieber 2010). Loss of gene function is often induced by premature stop codons downstream of the frameshift mutation using the CRISPR/Cas9 system approach

Multiplex CRISPR/Cas9 system impairs HCMV replication by excising an essential viral gene

anti-viral CRISPR/Cas9 strategies to target the UL122/123 gene, a key regulator of lytic replication and reactivation from latency. The singleplex strategy contains one gRNA to target the start codon ... viruses. It mainly induced large deletions (80±95%) after an exposure period of only eight days. Moreover, to reach this goal, high Cas9 expression is needed to target all copies of the viral genome before

indCAPS: A tool for designing screening primers for CRISPR/Cas9 mutagenesis events

digest can be used to distinguish the two alleles. However, in the case of most CRISPR-induced alleles, no such restriction sites are present in the target sequences. In this case, a derived CAPS (dCAPS ... the design of CAPS and dCAPS oligonucleotide primers used in PCR amplification of target sequences in order to screen for editing events following CRISPR/Cas9-mediated mutagenesis [ 13 ]. The tool is

One-step genetic correction of hemoglobin E/beta-thalassemia patient-derived iPSCs by the CRISPR/Cas9 system

CRISPR/Cas9 system to target the hemoglobin E mutation from one allele of the HBB gene by homology-directed repair with a single-stranded DNA oligonucleotide template. DNA sequences of the corrected iPSCs ... patients with HbE/ β-thalassemia in the future. Induced pluripotent stem cells; Thalassemia; Hematopoietic differentiation; Genetic correction; CRISPR/Cas9 Background HbE/β-thalassemia, a double

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attractive target for receptor-mediated imaging and therapy. Another interesting approach would be the use of GRPR radioligands for pre-operative imaging and subsequent radio-guided surgery, with the goal to ... improve surgical outcome. GRPR radioligands were successfully implemented in clinical studies, especially Sarabesin 3 (SB3) is an appealing GRPR antagonist with high receptor affinity. Gallium-68 labeled

Effect of <Emphasis Type=

. 2012) , the seed production potential of annual forbs is high in semi-arid rangelands due to their strong adaptation and survival strategies (Veenendaal et al. 1996), and this might lead to the ... herbaceous species, soil samples at a depth of 0-20 cm were taken for analyzing soil physical and chemical properties. Results: Highly palatable grasses and legumes based on the frequency of occurrence were

Improvement of <Emphasis Type=

fermentation using Escherichia coli or Corynebacterium glutamicum. In C. glutamicum, development of high l-ornithine producing C. glutamicum was achieved by deletion of argF, but was accompanied by growth ... genetic engineering for developing high target-compound producing strains. However, deletion of growth-coupled genes can induce some undesired results, such as a growth deficiency or extra nutritional