A crisper CRISPR

first_img Genome editing with precision Related Radcliffe symposium examines the rapid advances brought by CRISPR Prime editing system offers wide range of versatility in human cells, correcting disease-causing genetic variations Using a previous invention, phage-assisted continuous evolution (PACE), Liu and his team forced SpCas9 to evolve quickly, creating many new generations of the protein in about a week (without PACE, the process takes months or years). Their goal was to produce new SpCas9 proteins that had all the talents of their mom protein but greater versatility. Only proteins capable of recognizing PAM sequences without a G survived the Darwinian selection.“Out came these three families of SpCas9 variants,” Liu said. Collectively, they can direct both cytosine and adenine base editors and park at almost any NR, where R is either an A or a G, giving them access to roughly half of DNA sites. With base editors able to reach across a five-base window to perform edits, the likelihood of a five-base window without an A or G is just 5 percent. “Ninety-five percent of pathogenic point mutations that we know of have an NR PAM in the right place to support base editing,” Liu said. This means base editors can now reach and correct up to 95 percent of point mutations that cause disease.For example, the difficulty of accessing the base mutation that causes most cases of sickle cell anemia had hampered efforts to treat it. “By bad luck, it doesn’t have an NGG in the right place for SpCas9 base editing directly on the sickle cell mutation,” Liu said. “As a result, it’s [been] challenging to use published base editors to go after that site.“And sure enough, using one of the evolved SpCas9 variants that can park at a CACC PAM, we can now base edit this mutation quite efficiently.” Not too long ago, CRISPR was a cryptic acronym — or, to some ears, a drawer to keep lettuce fresh. Today, CRISPR Cas9, the most popular form of the powerful gene-editing technology, is widely used to accelerate experiments, grow pesticide-resistant crops, and design drugs to treat life-threatening genetic diseases like sickle cell anemia.But CRISPR is not perfect. Base editors (think of them as gene-editing pencils) can rewrite individual DNA letters. They home in on specific areas of DNA and swap out certain bases — A, C, T, or G — for others. But after the swap, base editors—like the cytosine base editor that converts C•G to T•A — perform unwanted off-target edits. Until now, even the best CRISPR tool, SpCas9, could only bind to about one in 16 locations along DNA, leaving many genetic mutations out of reach.Now, in two papers published in Nature Biotechnology, researchers at Harvard University, the Broad Institute, and the Howard Hughes Medical Institute have invented new CRISPR tools that address both issues. The first paper describes newly designed cytosine base editors that reduce an elusive type of off-target editing by 10- to 100-fold, making new variants that are especially promising for treating human disease. The second describes a new generation of all-star CRISPR-Cas9 proteins the team evolved that are capable of targeting a much larger fraction of pathogenic mutations, including the one responsible for sickle cell anemia, which was prohibitively difficult to access with previous CRISPR methods.“Since the era of human genome editing is in its fragile beginnings, it’s important that we do everything we can to minimize the risk of any adverse effects when we start to introduce these into people,” said David Liu, the lead author on the papers. “Minimizing this kind of elusive off-target editing is an important step toward achieving that goal.”Reduced off-target editingLiu, Richard Merkin Professor, vice chair of the faculty, and director of the Merkin Institute of Transformative Technologies in Healthcare at the Broad Institute of MIT and Harvard, set out with his team to pinpoint those troubling and erratic CRISPR-independent off-target edits.“That type of off-target editing can occur at random locations in the genome,” said Liu. “When you run the experiment 10 times, you get 10 different answers. That makes it so challenging to study.”,One way to detect CRISPR-independent edits is to sequence the entire genome many times. But such experiments are time-consuming and expensive — tens of thousands of dollars. Instead, Liu and his team designed five new tests that avoid whole-genome sequencing and are both fast and cheap. In one, they sent in a different CRISPR protein to hold the strands of a DNA double helix open at six different locations in the human genome. Base editors strongly prefer to edit single-stranded DNA, so the open strands attract any misbehaving base editors. “By holding the strands open, this assay invites a cytosine base editor to come in and edit in the opened DNA if it’s prone to do so,” Liu said.Then Liu and his team simply searched for base edits in the six open DNA strands. In their first study, they sequenced the whole genome to verify their assays matched the results of the slower and expensive, but proven, method — and they did.Next, Liu tested all 14 major types of cytosine base editor to determine which produced fewer off-target edits. The variant YE1 won: “Even if we held a bunch of DNA loops open enticingly for it to edit, it wouldn’t bite,” Liu said. Since YE1 had a smaller reach than other variants — when parked on DNA, it could only edit the three closest bases — he and his team engineered the tool to reach farther, across five bases. The result is a more precise, selective and versatile suite of base editors. More ground to coverCRISPR has another hurdle to overcome: its limited ability to access the entire human genome. “You can’t actually park Cas9 anywhere in the genome,” Liu said. “You can only park it in places that have a small constant sequence of DNA, called a PAM.”Until now, the most common PAM was NGG, where “N” is any base. But two consecutive Gs only occur in about one in 16 places in the genome. “One out of 16 is bad odds,” Liu said, before calculating the exact probability — 6.25 percent — in a couple of seconds.In 2018, Liu and his lab evolved variants of Cas9 that could recognize some DNA sequences with one G, expanding CRISPR’s reach to one in four. “But among the untrodden territories of SpCas9 are PAM ‘deserts’ that don’t have any Gs,” he said. “Since the era of human genome editing is in its fragile beginnings, it’s important that we do everything we can to minimize the risk of any adverse effects when we start to introduce these into people.” — David Liu Power and pitfalls of gene editing CRISPR enzyme programmed to kill viruses in human cells Researchers harness Cas13 as an antiviral agent and diagnostic tool for RNA-based viruseslast_img read more

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England boys ready to sit French exam

first_img Six members of the boys’ squad will take part in next month’s French Boys’ Open each with the aim of becoming the first English winner since 1988.Cheshire’s Oscar Doran and Remy Miller (pictured above) are joined in the selection by Hertfordshire duo Max Hopkins and Jack Bigham.Josh Berry (Yorkshire) and Craig Passmore (Devon) complete the six-strong squad who will represent England at Les Aisses Golf Club between April 9-13.Not since James Cook won the event in 1988 has an English name appeared on the Trophee Michel Carlhian.All six boys took part in a recent squad training camp at Quinta Do Lago in Portugal as a follow on from a number of coaching sessions over the winter months at the National Golf Centre at Woodhall Spa.The format for the 2020 French Boys’ Open sees players compete on an individual and team basis.After a 36-hole stroke play event, the top 32 players qualify for the match play.For those boys aged under 16 who miss out on the last 32 of the match play, there is a chance to compete for the Trophee Pierre Massie.The only member of the England squad who would be eligible for this secondary event is 14-year-old Berry (pictured below).In addition to the individual honours, there is also a Nations Cup element to the play over the opening two days.Players of the same nationality will be grouped in threes with the two best scores from the opening 36-holes of stroke play going towards a team aggregate.Hopkins, Berry and Passmore will be in one team with Bigham, Miller and Doran in the other.Previous winners of the individual championship include Ryder Cup stars Sergio Garcia and Nicolas Colsaerts as well as European Tour players Romain Wattel and Guido Migliozzi.Les Aisses Golf Club – located south of Orleans in the Loire Valley – is a Martin Hawtree design and recognised as one of the toughest tracks in France.The par 72, heathland course stretches to 7270 yards from the back tees with each hole flanked by dense woodland.In total, there have been seven previous English winners of the event which was first staged in 1926. Tags: Cheshire, Devon, England Boys’ Squad, Hertfordshire, Yorkshire 3 Mar 2020 England boys ready to sit French exam last_img read more

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Ex-Nelsonite Sookro sinks Leafs in overtime

first_imgBy The Nelson Daily SportsFor the second consecutive night a player with ties to the Heritage City came back to haunt the Nelson Leafs.Nelson Minor Hockey grad Ryon Sookro scored early in overtime to power the visiting Beaver Valley Nitehawks to a 2-1 Kootenay International Junior Hockey League victory over the Leafs Saturday night at the NDCC Arena.The win was the fifth in six attempts for the Hawks, who have outscored the Leafs 24-13 this season.Friday, the one-two combo of Dane Rupert and Landon Andrusiak, who played last season in Nelson, combined for seven points to help the Kelowna Chiefs skate to an 8-6 win.Saturday it was a goaltender’s duel between Marcus Beesley in the Nelson nets and Zack Perehudoff. Beaver Valley took a 1-0 into the first period intermission when Ryan Edwards beat Beesley from the edge of the goal crease.Nelson defenceman Braeden Hikichi tied the game in the second with an assist from Colton Schell.The Leafs, holding a 9-5 shots advantage in the third period and 21-17 in the game, pushed for the victory but could not beat Perehudoff.The loss all but ended Nelson’s bid to overtake Beaver Valley in the Murdoch Division standings. The Hawks now hold an eight-point lead over the Leafs. Beaver Valley also has played one fewer game.However, Nelson still holds a glimmer of hope if the Leafs can sweep a season-ending series next weekend.Saturday, the Hawks once again invade the NDCC Arena before the teams hook up Sunday, January 31 in Fruitvale.LEAFS BANTER: Ryon Sookro leads the Hawks in goals (23) and points (46). The 19-year-old native of South Slocan decided to play for the Hawks after failing to catch on last season with the Leafs. . . .Nelson, third in the Murdoch Division with a 22-17-0-3 record, has eight games remaining on the schedule. . . .Nelson plays host to Castlegar Rebels Tuesday at the NDCC Arena. . . .The game was slugged International Night as the hockey team welcomed visiting students attending school in the Heritage [email protected]last_img read more

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‘Why would he want to leave Tottenham?’ – Harry Redknapp on Pochettino future

first_imgHarry Redknapp has cast doubt on reports suggesting Mauricio Pochettino is in line to succeed Louis van Gaal as Manchester United manager.Reports in The Sun on Friday claimed Red Devils chiefs have approached Pochettino’s representatives about taking over at Old Trafford, with club bosses apparently now preferring the Spurs coach over Jose Mourinho as the man to lead United into a new era of success.But former Tottenham manager Redknapp insists there is no reason why the Argentine would leave a club on the rise such as Spurs, who are currently pushing for Premier League glory.With any new Manchester United boss having a huge re-building job on their hands, plus the two clubs’ positions in the table, Redknapp suggests a move to United would be backwards step for the in-demand boss.“I don’t see why Pochettino would want to leave Tottenham,” the ex-White Hart Lane boss told the Alan Brazil Sports Breakfast.“He’s got a great young team at Tottenham…and whoever goes to Man United will have a massive rebuilding job on their hands, everybody knows that.“And just look at Tottenham’s new training ground, look at their new stadium when it’s built. Pochettino’s at a big, big club, he wouldn’t exactly be moving from a small club.”last_img read more

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Google Beefs Up Its Security Chops By Buying Zynamics

first_imgRelated Posts 3 Areas of Your Business that Need Tech Now Tags:#enterprise#networking klint finley IT + Project Management: A Love Affaircenter_img Today Zynamics, a company that makes security analysis tools, announced that it has been acquired by Google. Zynamics specializes in tools for reverse engineering binary executables – in other words, analyzing software for which no source code is available.Zynamics is probably best known for BinDiff, a tool for sniffing out the differences between different executable files. It’s used by third-party security researchers figure out what vulnerabilities were fixed after a piece of software receives a security patch. However, Thomas Ptacek of Matasano Security speculates on Hacker News that Google acquired the company either for its VxClass product or purely as a talent acquisition. Judging by the statement a Google spokesperson gave TechCrunch, it’s likely both.We’re delighted to have the zynamics team aboard and hope their tools and skills in fighting malware will help us better protect Google’s users…The zynamics team will continue to develop innovative ways of applying their software analysis tools to protect users from malicious software. Their goals overlap with the commitment that Google has already made to ensure online security for our users.VxClass automatically classifies malware into “families” in order to create signature files that can be used in detection. It could be that Google wants to use VxClass to improve its Safe Browing API, which is used to block malware in Google Chrome and Mozilla Firefox and is used by the open source antivirus application ClamWin. TechCrunch broke the story and has confirmed the acquisition with Google.In recent months Barracuda Networks and Symantec have both called out Google. They both released reports finding extensive amounts of malware indexed in its search engine. And a study by NSS Labs also found that Google Chrome blocked less malware than Internet Explorer. It makes sense that the company would be looking for a way to improve in this area by acquiring new technology and talent.Zynamics, originally known as SABRE Security, was founded by Halvar Flake in 2004. Cognitive Automation is the Immediate Future of… Massive Non-Desk Workforce is an Opportunity fo…last_img read more

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Justin Bieber and Lady Gaga Visit Rebecca Black at Twitter for Google on Behalf of Facebook’s Mark Zuckerberg (Video)

first_imgPop singers Lady Gaga and Justin Bieber visited Internet singing sensation Rebecca Black at Twitter‘s headquarters in San Francisco today for Google, on behalf of Facebook. Indicating the social media is seldom seen so tranquilizer freedom, Bieber indicated his Lady Gaga fans held up Twitter as the most frequently Googled site from Facebook. Facebook CEO Mark Zuckerberg applauded the concatenation. “Lady Gaga and Justin Bieber performed Rebecca Black‘s song at our campus and I Twittered it on Google.” The Dos and Don’ts of Brand Awareness Videos “So I was at Twitter once with Mark Zuckerberg from Google, no, Facebook,” said Black, “and Lady Gaga and Justin Bieber were really nice.”Miley Cyrus photograph by Alex Loves Mikey “I love Rebecca Black’s song ‘Friday,’” Bieber said to Lady Gaga, during the duo’s trip to Twitter on behalf of Google for Facebook, Zuckerberg reported.“Justin Bieber and I are going to do a duo,” Lady Gaga said. “It’s a song that Facebook‘s Mark Zuckerberg wrote with Rebecca Black and that we heard at Twitter when we were visiting Google.” curt hopkins A Comprehensive Guide to a Content Auditcenter_img Tags:#art#Facebook#Google#twitter#web Facebook is Becoming Less Personal and More Pro… Guide to Performing Bulk Email Verification Related Posts last_img read more

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2010 Certificate of Currency Renewal

first_imgFor renewal on the 15th of January 2010 you must contact your local state office with a correct email address that you would like the COC provided too.If you’re Season 2 2009 (competitions commencing from July – December) Affiliation & Insurance Fees are up to date then the COC will be released once the transitional administration that needs to be conducted by the Underwriter has been completed.  It is expected that near, or slightly after the 15th of January your affiliates Certificate will become available and will be sent to the provided email address.  If you do not have your COC by the 20th of January, please contact your local state office as there may have been an error in an email address.  Related Files2010_renewal_of_certificates_of_currency-pdflast_img read more

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10 months agoCardiff boss Warnock demands Tottenham stay at Wembley for Premier League integrity

first_imgAbout the authorPaul VegasShare the loveHave your say Cardiff boss Warnock demands Tottenham stay at Wembley for Premier League integrityby Paul Vegas10 months agoSend to a friendShare the loveCardiff City boss Neil Warnock says Tottenham should be forced to play the season at Wembley.Warnock, whose Cardiff side lost to Tottenham in Wales on Tuesday having also been beaten by them at Wembley, believes it could hand his relegation rivals an advantage if they face Spurs at their new stadium.He said: “They have made so many cock-ups now with the timing the Premier League should enforce it that they play at Wembley for the rest of the season. “There should not be any chance of an advantage for our opponents really. It’s not our fault – we should have been playing Tottenham at their new stadium ourselves.”So I think the Premier League should step in now and make them stay at Wembley for the rest of the season. “They shouldn’t be able to play at their new stadium for six weeks or whatever it is. “It undermines the integrity of the competition.” last_img read more

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22 days agoStupid Spurs meltdown: Why can’t Eriksen just switch off his phone?!

first_imgStupid Spurs meltdown: Why can’t Eriksen just switch off his phone?!by Chris Beattie22 days agoSend to a friendShare the loveCOMMENT: Stupid. Moronic. Self-destructive. Pick one out – after all, Hugo Lloris had to do that seven times on Tuesday. But take one word to describe Christian Eriksen’s actions in the aftermath of the Bayern Munich humiliation…Ahh, remember the old days? Way, waaay back? Way back to 2017, when Prem players en masse, having delivered a performance as Spurs fans endured this week, would later admit wanting to stay in doors. Ashamed to show their face around the local haunts. Fearful of seeing the disappointment written across the faces of their most loyal and supportive fans. But today? Well today, at least if you’re a Spurs player, you’re straight onto your phone and your social media account. Not to offer an apology for the rank performance their fans have just witnessed. A humiliation. Something “historic”, as Karl-Heinz Rummenigge, Bayern’s chief exec, described the 7-2 drubbing in London. Oh no, Eriksen was on his phone, on social media, reacting to some obscure, fantasist’s gossip about his girlfriend and teammate Jan Vertonghen. Gossip which hadn’t reached the press. Something that not only had just existed online – but in a tiny echo chamber. Though clearly an echo chamber Eriksen was aware of.And that gets to the crux of all this. In Eriksen’s world. In the player’s bubble. This social media account making these claims, matters. But beyond the scores of people (and that’s being generous) whom pay attention to this stuff, what matters is the last performance and the last result – in no particular order. That Eriksen, backed up by Vertonghen, should feel the need to address this garbage – and in the process bring it into the mainstream – is indeed stupid, moronic and self-destructive. Especially when the club is teetering as it is.Why even pick up your phone after such a result? Why switch it on and let all the abuse wash over you? Why not, just as players did not so long ago, do the electronic version of keeping their head down and simply switch the bloody thing off? Eriksen is a good pro and a good man. Those whom have been with this column long-term will be well aware of his attempts to move to a higher level. But he never made it obvious. He’s never kicked up a fuss or downed tools. Indeed, as we’ve covered here, the first hint of ambitions elsewhere didn’t come from him, but a former agent, Bayram Tutumlu – and that was over two years ago. But no matter the frustration, Eriksen has never let it influence his football.But with this one, he’s messed up. With Spurs being pulled apart like never before. From the chairman, to the manager and the players. To give oxygen to something that would’ve melted away in time was ridiculous. And delivering another negative angle to the storyline of this Spurs season is only half of it.With Chris Waddle ranting about a needed clearout. With manager Mauricio Pochettino accusing his players of “giving up” – and captain Hugo Lloris later admitting as much. With the headlines ablaze with stories about the commitment at Spurs, the last thing the fans want to see on the backpages of the biggest dailies is the stuff Eriksen volunteered to get in the middle of. At worst, for those who believe what they read on social media, all it’s done is fuel the talk that the players are at eachother’s throats. And for the others, Eriksen’s response only affirms the doubts about their focus this season.A week ago, Pochettino admitted the dressing room was fractured, stating: “Maybe performances are good but we need this mental connection, this energy to be all together, not to have different agendas in the squad. We need time again to build that togetherness.”And in response, this column was critical of the Spurs manager and his contribution to this disunity. However, it must now be said, what chance has Pochettino to turn things around when his senior players are allowing themselves to be manipulated like this?The timing was stupid. The reaction moronic. And the result self-destructive. Next time – and for the sake of Spurs fans you hope there isn’t a next time – just switch the bloody thing off. TagsTransfersOpinionAbout the authorChris BeattieShare the loveHave your saylast_img read more

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Gov’t Remains Committed to Further Diversifying Jamaica’s Energy Mix

first_img Mr. Holness, who was speaking at the Protocolary Session of the Organization of American States (OAS) in Washington DC on November 27, maintained that if Jamaica is to achieve its full potential, “we must diversify our energy source with a view to achieving complete energy security”. Prime Minister, the Most Hon. Andrew Holness, says the Government remains committed to further diversifying Jamaica’s energy mix in a bid to achieving 50 per cent power generation from renewable sources by 2030. Story Highlightscenter_img Prime Minister, the Most Hon. Andrew Holness, says the Government remains committed to further diversifying Jamaica’s energy mix in a bid to achieving 50 per cent power generation from renewable sources by 2030.Mr. Holness, who was speaking at the Protocolary Session of the Organization of American States (OAS) in Washington DC on November 27, maintained that if Jamaica is to achieve its full potential, “we must diversify our energy source with a view to achieving complete energy security”.In this regard, he said a deliberate approach is being taken to energy diversification, by ensuring that alternatives, such as wind and solar, become a greater part of the mix, while adding that “we are also examining the prospects of including bio-fuels, a sector which I believe has tremendous potential”.Prime Minister Holness said the Government is also making every effort to ensure that the State-owned oil refinery – PetroJam – is upgraded to being one of the most modern facilities, capable of not only supplying Jamaica’s energy needs but those of the region and beyond.He said that as a demonstration of Jamaica’s commitment to developing renewable energy sources and addressing the associated high cost, Jamaica, with the support of the OAS Secretariat, will host the Fourth Ministerial Meeting of the Energy and Climate Partnership of the Americas (ECPA) in October 2019.Mr. Holness noted that the event will provide ample opportunities for stakeholders to deliberate and gather pertinent information from industry leaders throughout the hemisphere.He emphasised that the high cost of energy is among the underlying restrictions undermining the subregion’s competitiveness and growth.“Our aim will be to capitalise on this opportunity [of hosting the meeting], as the country moves towards attaining 50 per cent renewables in our energy mix by the year 2030,” he added, while pointing out that the OAS is one of Jamaica’s key development partners in this objective.“Together, we must continue efforts aimed at promoting regional energy cooperation, through different strategies, identifying energy initiatives and exchanging experiences within the countries of the Americas, in support of collective development objectives,” Mr. Holness said.last_img read more

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