Sentences with phrase «of nucleases»

Remarkably, some prokaryotes employ a structurally distinct family of nucleases with a dual function e.g., in DNA repair and antiviral immunity.

Currently, Cas - Analyzer supports a variety of nucleases, including single nucleases (SpCas9, StCas9, NmCas9, SaCas9, CjCas9, and AsCpf1 / LbCpf1) and paired nucleases (ZFNs, TALENs, Cas9 nickases, and dCas9 - FokI nucleases).

Genetically - modified mouse models: pros and cons of nucleases - based technologies.

To go further in the TREX2 role in homeostasis and skin pathogenesis, the researchers wanted to see the role of this nuclease on psoriasis.

A stopped - flow spectrofluorometer was used to study the kinetics renaturation of nuclease from the acidified form on neutralization, the refolding is fast and the data can be described as a sequence of two first - order processes with half times of about 55 and 350 milliseconds, respectively.

The point mutation was induced by forming a synthetic complex through removal of nuclease activity from the CRISPR system — a technique using artificial nuclease — and addition of deaminase, a deaminizing (base - modifying) enzyme, and then expressing it in yeasts and mammalian cells.

The treated PCR products were diluted with 50 uL of nuclease - free water before Quant - IT ™ PicoGreen ® dsDNA Assay Kit quantification (http://www.lifetechnologies.com) on a plate reader.

The 50 uL PCR reaction consisted of the following mixture: 10 uL 5X LongAmp Taq Reaction Buffer, 1.5 uL of dNTPs (1 mM), 4 uL of each primer pairs (10 uM each, see below for details), 2 uL of template DNA, 2 uL of LongAmp Hot Start Taq DNA polymerase (2500 U / mL) and 30.5 uL of nuclease - free water.

H. Efficient CRISPR / Cas9 - Mediated Genome Editing in Mice by Zygote Electroporation of Nuclease.

In MMEJ pathway, we achieve efficient gene disruption in human cell lines and animals by developing a computer program that assists the choice of nuclease target sites based on microhomology prediction.

These systems, however, require efficient design and time - consuming assembly of nuclease constructs for DNA targeting.

gRNAs were co-transfected with reporter, dCas9 - VPR, a tripartite transcriptional activator fused to the C - terminus of nuclease - null Streptococcus pyogenes Cas9, and an EBFP2 expressing control plasmid into HEK293T cells.

Not exact matches

Announced a worldwide collaboration with Sangamo Therapeutics, Inc. (Sangamo) using Sangamo's zinc finger nuclease technology platform for the development of next - generation ex vivo cell therapies in oncology.

These risks and uncertainties include: Gilead's ability to achieve its anticipated full year 2018 financial results; Gilead's ability to sustain growth in revenues for its antiviral and other programs; the risk that private and public payers may be reluctant to provide, or continue to provide, coverage or reimbursement for new products, including Vosevi, Yescarta, Epclusa, Harvoni, Genvoya, Odefsey, Descovy, Biktarvy and Vemlidy ®; austerity measures in European countries that may increase the amount of discount required on Gilead's products; an increase in discounts, chargebacks and rebates due to ongoing contracts and future negotiations with commercial and government payers; a larger than anticipated shift in payer mix to more highly discounted payer segments and geographic regions and decreases in treatment duration; availability of funding for state AIDS Drug Assistance Programs (ADAPs); continued fluctuations in ADAP purchases driven by federal and state grant cycles which may not mirror patient demand and may cause fluctuations in Gilead's earnings; market share and price erosion caused by the introduction of generic versions of Viread and Truvada, an uncertain global macroeconomic environment; and potential amendments to the Affordable Care Act or other government action that could have the effect of lowering prices or reducing the number of insured patients; the possibility of unfavorable results from clinical trials involving investigational compounds; Gilead's ability to initiate clinical trials in its currently anticipated timeframes; the levels of inventory held by wholesalers and retailers which may cause fluctuations in Gilead's earnings; Kite's ability to develop and commercialize cell therapies utilizing the zinc finger nuclease technology platform and realize the benefits of the Sangamo partnership; Gilead's ability to submit new drug applications for new product candidates in the timelines currently anticipated; Gilead's ability to receive regulatory approvals in a timely manner or at all, for new and current products, including Biktarvy; Gilead's ability to successfully commercialize its products, including Biktarvy; the risk that physicians and patients may not see advantages of these products over other therapies and may therefore be reluctant to prescribe the products; Gilead's ability to successfully develop its hematology / oncology and inflammation / respiratory programs; safety and efficacy data from clinical studies may not warrant further development of Gilead's product candidates, including GS - 9620 and Yescarta in combination with Pfizer's utomilumab; Gilead's ability to pay dividends or complete its share repurchase program due to changes in its stock price, corporate or other market conditions; fluctuations in the foreign exchange rate of the U.S. dollar that may cause an unfavorable foreign currency exchange impact on Gilead's future revenues and pre-tax earnings; and other risks identified from time to time in Gilead's reports filed with the U.S. Securities and Exchange Commission (the SEC).

In recent years several techniques, such as CRISPR / Cas9 or zinc finger nucleases have been experimented to directly modify the DNA of plants and animals.

«CRISPR has proven so easy and inexpensive that Dana Carroll of the University of Utah, Salt Lake City, who spearheaded the development of zinc finger nucleases, [one of its competitors,] says it has brought about the «democratization of gene targeting.

By using engineered zinc - finger nucleases (ZFNs) designed to target an integrated reporter and two endogenous rat genes, Immunoglobulin M (IgM) and Rab38, we demonstrate that a single injection of DNA or messenger RNA encoding ZFNs into the one - cell rat embryo leads to a high frequency of animals carrying 25 to 100 % disruption at the target locus.

Scientists can select from a variety of scalpels, including zinc finger nucleases, TALENs and CRISPR / Cas9.

In plants and Caenorhabditis elegans, two distinct populations of small RNAs have been proposed to participate in RNAi: «Primary siRNAs» (derived from DICER nuclease - mediated cleavage of the original trigger) and «secondary siRNAs» [additional small RNAs whose synthesis requires an RNA - directed RNA polymerase (RdRP)-RSB-.

Nuclease and chemical protection studies with the 52mer helped to define the DNA base pairs that contributed to the specificity of binding.

GUIDE - seq enables genome - wide profiling of off - target cleavage by CRISPR - Cas nucleases.

Development and evaluation of a fluorogenic 5 ′ nuclease assay to detect and differentiate between Ebola virus subtypes Zaire and Sudan

Talk of curing AIDS made front - page news last year, in part due to an astonishing new gene - editing technology: lab - engineered proteins called zinc finger nucleases.

And in another advance, virologist Paula Cannon of the University of Southern California used zinc finger nucleases to create human stem cells that lack CCR5.

KIM Jin - soo, Director of the IBS Center for Genome Editing and corresponding author of the two studies commented, «Since the two studies have proved the superior specificity of Cpf1, this new nuclease will be more widely used for precise genome editing that does not produce any unintended mutations.

Furthermore, by adding guanine nucleotides at the end of sgRNA (single guided RNA) that composes CRISPR - Cas9, they have successfully created this highly - developed programmable nuclease, which has no measurable off - target effects in the human genome.

But CRISPR has proven so easy and inexpensive that Dana Carroll of the University of Utah, Salt Lake City, who spearheaded the development of zinc finger nucleases, says it has brought about the «democratization of gene targeting.»

The viral scraps serve as an infection memory bank: From them, bacteria create guide RNAs that can seek out the DNA of returning viruses before chopping up the viral genes with a nuclease.

Shown to work just 3 years ago, CRISPR consists of a an enzyme called a nuclease and a piece of RNA that homes in on a targeted DNA sequence, enabling the enzyme to introduce precisely targeted mutations, corrections to mutations, or other alterations.

Just this week, for example, a team led by Feng Zhang of the Broad Institute of Harvard and MIT, one of the pioneers of the method, published a paper in Science on engineering the nuclease part of CRISPR so that it more accurately cuts the intended DNA target.

Meanwhile, Cellectis announced it now has «an umbrella patent» that its CEO, Andre Choulika, says «covers most of the gene editing procedures done with a nuclease,» including those based on CRISPR - Cas 9, TALENs, zinc fingers, and many meganucleases.

A similar mutation, detected by S1 nuclease mapping of LDL receptor messenger RNA, occurred in a patient with familial hypercholesterolemia whose receptor also fails to be transported to the cell surface.

Conventional CRISPR uses a guide RNA (gRNA) coupled with an enzyme known as a nuclease, most commonly Cas9, that together attach to a specific stretch of DNA bases; the nuclease then snips the double helix.

«Gene editing based on nucleases is very good at inactivating genes,» says CRISPR researcher Feng Zhang of the Broad Institute in Cambridge, Massachusetts.

What's more, say the researchers, the cancer - causing effects of off - target deletions mistakenly created by the V (D) J enzyme need to be considered in designing site - specific enzymes for genome modification such as zinc - finger nucleases, TALENS, or CRISPRs.

According to Director Jin - Soo Kim, «We used CRISPR RGENs [RNA - guided engineered nucleases] to repair two recurrent, large chromosomal inversions responsible for almost half of all severe hemophilia A cases.»

McNamara acknowledges previous research by Arthur Arnone, UI professor emeritus in biochemistry, who was the first to define the structure of the S. aureus nuclease.

The Mfd protein was shown to (i) displace RNAP stalled at a lesion in an adenosine triphosphate - dependent reaction, (ii) bind to the damage recognition subunit (UvrA) of the excision nuclease, and (iii) stimulate the repair of the transcribed strand only when transcription is taking place.

«Research team evolves CRISPR - Cas9 nucleases with novel properties: Engineered variants of gene - cutting enzyme double the targeting range, reduce off - target effects.»

The class 2 systems have evolved from class 1 systems via the insertion of transposable elements encoding various nucleases, and are now being used as tools for genome editing.

A team of Massachusetts General Hospital (MGH) researchers has found a way to expand the use and precision of the powerful gene - editing tools called CRISPR - Cas9 RNA - guided nucleases.

Last year, researchers targeted and destroyed this gene in the T - cells of 12 people with HIV using custom - made proteins called zinc finger nucleases.

In an effort to combine the respective advantages of the AAV and CRISPR - based approaches, several groups have recently worked to enhance the efficiency of AAV - based gene editing via the introduction of a double strand break by using a targeted nuclease.

Together the stable disruption of CXCR4 as determined by both the surveyor nuclease assay and flow cytometry suggests that CXCR4 disruption did not negatively impact cell viability or growth in humanized NSG mice over a two - month period.

The trial is using a form of DNA scissors called zinc finger nucleases (ZFNs).

As an approach to inactivating CCR5, we introduced CCR5 - specific zinc - finger nucleases into human CD4 + T cells prior to adoptive transfer, but the need to protect cells from virus strains that use CXCR4 (X4) in place of or in addition to CCR5 (R5X4) remains.

Cell growth was monitored every 48 hours post-stimulation for approximately two weeks and the efficiency of CXCR4 disruption was assessed at day five post-transduction by both the Surveyor nuclease assay and by deep - sequencing of the CXCR4 target site.

Samples of cDNA were diluted 1:25 in nuclease - free water (Qiagen Inc.).

In the presence of Bk132, treatment with X4 - ZFNs conferred protection at 14 d.p.i (p =.05); however, this protection wanes by 34 d.p.i. (p =.88)(B) Cxcr4 disruption frequency was assessed by the surveyor nuclease assay in both peripheral blood (p <.001) and spleen (p <.001).

Here we describe engineering a pair of zinc finger nucleases that, when introduced into human T cells, efficiently disrupt cxcr4 by cleavage and error - prone non-homologous DNA end - joining.