Advanced search    

Search: Potential high frequency target mutagenesis induced CRISPR Cas9 Emphasis Type

The page you are looking for is no longer available: Potential high-frequency off-target mutagenesis induced by CRISPR/Cas9 in <Emphasis Type=. Showing search results instead.

1,497,358 papers found. Showing first 1,000 results.
Use AND, OR, NOT, +word, -word, "long phrase", (parentheses) to fine-tune your search.

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

Protein Engineering Strategies to Expand CRISPR-Cas9 Applications

complex that can be reprogrammed to specifically interact with a desired DNA sequence target. CRISPR-Cas9 has been used in a wide variety of applications ranging from basic science to the clinic, such as ... current strategies for engineering SpCas9 through end-to-end fusion and domain insertion [43]. 2.1. Reducing Off-Target Events One of the main limitations of the CRISPR-Cas9 system is the high level of off

Repair of double-strand breaks induced by CRISPRCas9 leads to large deletions and complex rearrangements

target site and distal off-target sequences, leading to the conclusion that CRISPRCas9 was reasonably specific. Here we report significant on-target mutagenesis, such as large deletions and more complex ... . et al. Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nat. Biotechnol. 32, 569–576 (2014). CASPubMedArticleGoogle Scholar11. Fu, Y. et al. High-frequency off-target

High-fidelity CRISPR/Cas9- based gene-specific hydroxymethylation rescues gene expression and attenuates renal fibrosis

therapy. TET enzymes catalyze hydroxymethylation of methylated DNA, reactivating gene expression. We here report generation of a high-fidelity CRISPR/Cas9-based gene-specific dioxygenase by fusing an ... the mutated wild-type SpCas9 in this technology results in substantial off-target effects we developed a new high-fidelity Cas9-based approach which reduced off-target genes by 85%. The relevance of

Novel Epigenetic Techniques Provided by the CRISPR/Cas9 System

phenotype changes and cell identity will pave the way for us to understand diverse biological processes. In recent years, the emergence of CRISPR/Cas9 technology has provided us with new routes to the ... the same time, CRISPR/Cas9 could be a scalable and multiplex system for miRNA family mutagenesis. The mutant rate could be as high as 90% [56]. Moreover, the CRISPR/Cas9 deletion-based method could be

CRISPR/Cas9-Mediated Generation of Guangxi Bama Minipigs Harboring Three Mutations in α-Synuclein Causing Parkinson’s Disease

as an ideal model for PD research. In the present study, we attempted to model PD by generating Guangxi Bama minipigs with three PD-causing missense mutations (E46K, H50Q and G51D) in SCNA using CRISPR ... , specifically the CRISPR/Cas9 plasmid and the repair vector, were integrated into cell colonies 54# and 67# (see Supplementary Fig. S2). Off-target effect is another concern when CRISPR/Cas9 is used for cellular

Small molecule inhibitors and CRISPR/Cas9 mutagenesis demonstrate that SMYD2 and SMYD3 activity are dispensable for autonomous cancer cell proliferation

methyltransferases, SMYD2 and SMYD3. We show that tool compounds as well as CRISPR/Cas9 fail to reproduce many of the cell proliferation findings associated with SMYD2 and SMYD3 inhibition previously obtained with ... /Cas9 pooled screening has potential advantages over RNAi-based methods as it may have fewer off-target effects and leads to gene knockout as opposed to only gene knockdown. This may be especially

Engineering of Primary Human B cells with CRISPR/Cas9 Targeted Nuclease

, we report optimized CRISPR/Cas9 based genome engineering of primary human B cells. Our procedure involves enrichment of CD19+ B cells from PBMCs followed by activation, expansion, and electroporation ... . CRISPR/Cas9 RNPs can be used to induce chromosomal translocations One potential application of this technology is the induction of chromosomal translocations in a site-specific manner for the study of B

<Emphasis Type=

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

5′ capped and 3′ polyA-tailed sgRNAs enhance the efficiency of CRISPR-Cas9 system

adaptive immune system in a variety of bacteria and archaea (Terns and Terns, 2011) . The most commonly used Streptococcus pyogenes type II CRISPR-Cas9 system consists of Cas9 nuclease and two short RNAs ... CTsgRNA caused higher mutation rates at other off-target sites. When sgRNAs with different structures and Cas9 mRNA were delivered into human primary T cells, genome editing induced by unmodified AAVS1

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

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

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

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

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

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

<Emphasis Type=

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

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

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