Navigation Links
Rapid method of assembling new gene-editing tool could revolutionize genetic research
Date:4/9/2012

Development of a new way to make a powerful tool for altering gene sequences should greatly increase the ability of researchers to knock out or otherwise alter the expression of any gene they are studying. The new method allows investigators to quickly create a large number of TALENs (transcription activator-like effector nucleases), enzymes that target specific DNA sequences and have several advantages over zinc-finger nucleases (ZFNs), which have become a critical tool for investigating gene function and potential gene therapy applications.

"I believe that TALENs and the ability to make them in high throughput, which this new technology allows, could literally change the way much of biology is practiced by enabling rapid and simple targeted knockout of any gene of interest by any researcher," says J. Keith Joung, MD, PhD, associate chief for Research in the Massachusetts General Hospital (MGH) Department of Pathology and co-senior author of the report that will appear in Nature Biotechnology and has received advance online release.

TALENs take advantage of TAL effectors, proteins naturally secreted by a plant bacteria that are able to recognize specific base pairs of DNA. A string of the appropriate TAL effectors can be designed to recognize and bind to any desired DNA sequence. TALENs are created by attaching a nuclease, an enzyme that snips through both DNA strands at the desired location, allowing the introduction of new genetic material. TALENs are able to target longer gene sequences than is possible with ZFNs and are significantly easier to construct. But until now there has been no inexpensive, publicly available method of rapidly generating a large number of TALENs.

The method developed by Joung and his colleagues called the FLASH (fast ligation-based automatable solid-phase high-throughput) system assembles DNA fragments encoding a TALEN on a magnetic bead held in place by an external magnet, allowing automated construction by a liquid-handling robot of DNA that encodes as many as 96 TALENs in a single day at a cost of around $75 per TALEN. Joung's team also developed a manual version of FLASH that would allow labs without access to robotic equipment to construct up to 24 TALEN sequences a day. In their test of the system in human cells, the investigators found that FLASH-assembled TALENs were able to successfully induce breaks in 84 of 96 targeted genes known to be involved in cancer or in epigenetic regulation.

"Finding that 85 to 90 percent of FLASH-assembled TALENs have very high genome-editing activity in human cells means that we can essentially target any DNA sequence of interest, a capability that greatly exceeds what has been possible with other nucleases," says Jeffry D. Sander, PhD, co-senior author of the FLASH report and a fellow in Joung's laboratory. "The ability to make a TALEN for any DNA sequence with a high probability of success changes the way we think about gene-altering technology because now the question isn't whether you can target your gene of interest but rather which genes do you want to target and alter."

The research team also found that the longer a TALEN was, the less likely it was to have toxic effects on a cell, which they suspect may indicate that shorter TALENs have a greater probability of binding to and altering unintended gene sites. Joung notes that this supports the importance of designing longer TALENs for future research and potential therapeutic applications.

In 2008, Joung and colleagues at other institutions established the Zinc Finger Consortium (http://zincfingers.org), which has made a method of engineering ZFNs broadly available to academic laboratories. His team is now making the information and materials required to create TALENs with FLASH available within the academic community, and information about accessing those tools is available at http://TALengineering.org. Gene editing nucleases, including both ZFNs and TALENs, were recently named "Method of the Year" for 2011 by the journal Nature Methods.

Joung says, "While I believe that TALENs ease of design and better targeting range will probably make them a preferred option over ZFNs made by publicly available methods, ZFNs' smaller size and the less repetitive nature of their amino acid sequences may give them advantages for certain applications. For the time being, it will be important to continue developing both technologies." Joung is an associate professor of Pathology and Sander an instructor in Pathology at Harvard Medical School,


'/>"/>
Contact: Sue McGreevey
smcgreevey@partners.org
617-724-2764
Massachusetts General Hospital
Source:Eurekalert

Related biology news :

1. Reproducing early and often is the key to rapid evolution in plants
2. Forsyth scientist receives major grant to support rapid, accurate, affordable test for tuberculosis
3. Columbia University scientist devises new way to more rapidly generate bone tissue
4. Scattered light rapidly detects tumor response to chemotherapy
5. Study finds hemlock trees dying rapidly, affecting forest carbon cycle
6. HudsonAlpha investigator develops rapid response swine flu test
7. Rapid approach to identify influenza A virus mutations and drug resistance developed
8. German Research Minister Schavan: Rapid knowledge transfer can safe lives
9. K-State host to workshop on rapid methods to detect microorganisms in food
10. Growing Demand for Rapid Screening and Detection Systems Pushes the European Maritime Security Market, Finds Frost & Sullivan
11. Trial of new treatment for advanced melanoma shows rapid shrinking of tumors
Post Your Comments:
*Name:
*Comment:
*Email:
(Date:1/6/2017)... 6, 2017  Privately-held CalciMedica, Inc., announced that ... volunteers of a novel calcium release-activated calcium (CRAC) ... Acute pancreatitis, sudden painful inflammation ... but can be very serious.  In severe cases it ... extended hospital stays, time in the ICU and ...
(Date:1/6/2017)... 5, 2017  Delta ID Inc., a leader in ... for automotive at CES® 2017. Delta ID has collaborated ... demonstrate the use of iris scanning as a secure, ... driver in a car, and as a way to ... Delta ID and Gentex will demonstrate (booth ...
(Date:1/4/2017)... VEGAS , Jan. 4, 2017  CES ... performance biometric sensor technology, today announced the launch ... sensor systems, the highly-accurate biometric sensor modules ... biometric technology, experience and expertise. The two ... Benchmark designed specifically for hearables, and Benchmark BW2.0, ...
Breaking Biology News(10 mins):
(Date:1/12/2017)... ... 12, 2017 , ... Huffman Engineering, Inc. , a leader in control ... the company’s Lincoln office as a chemical engineer. In his new role, Beck ... in the life science manufacturing and water/wastewater industries. , Prior to joining Huffman Engineering, ...
(Date:1/12/2017)... 2017 The report "Direct-Fed Microbials Market by Type (Lactic Acid Bacteria ... and Region - Global Forecast to 2022", published by MarketsandMarkets, the global market ... to reach USD 1,399.6 Million by 2022, at a CAGR of 6.96% from ... ... MarketsandMarkets Logo ...
(Date:1/11/2017)... BELLINGHAM, Washington, USA (PRWEB) , ... January 11, ... ... SPIE, the international society for optics and photonics , are commending the ... and photonics, following the signing Friday by the President of the American Innovation ...
(Date:1/11/2017)... ... January 11, 2017 , ... IsoPlexis ... response analysis platform to measure the proteomic function of individual cells in patients, ... Research (SBIR) grant from the National Institute on Aging of the National Institutes ...
Breaking Biology Technology: