Major breakthrough! Developed based on the CRISPR one-time multiplex nucleic acid detection platform

Source: a major breakthrough! Developed based on the CRISPR one-time multiplex nucleic acid detection platform

2018 2 November 19/bio Valley BIOON/—in a new study, before the first time developed a rapid and inexpensive highly sensitive based on the CRISPR diagnostic tool—is called SHERLOCK（Specific High Sensitivity Enzymatic Reporter UnLOCKing the—the researchers greatly enhances this tool’s capabilities, and developed a micro-strip test method, this test method allows the naked eye to the test results, without the use of expensive equipment. Relevant research results in 2018 2 May 15, published online in the Science journal, The paper entitled“Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6 is”.

the researchers developed a simple test strip is used to display a single genetic marker test results, just like in a pregnancy test common visual cues. When the test strip is immersed in the treated sample after, there will be a line to indicate the target molecule is to be detected.

this new feature helps for field test, such as in epidemic outbreaks during the pave the way. These researchers also increased the SHERLOCK sensitivity, and increase the accurately determined quantitatively in the sample a target molecule levels and test a variety of target molecules of capacity. In short, these developments accelerate the SHERLOCK to quickly and accurately detect the sample of genetic characteristics, including pathogens and tumor DNA.

as this new Institute described above, these new developments build on these researchers to develop the SHERLOCK early version of the above, and there are more and more research the CRISPR system for genome editing with the use. This new study by the Massachusetts Institute of technology, Harvard University and the Broadbent Institute, the researchers led by the potential for research and global public health to produce a transformative impact.

papers communication the author, the Broadbent Institute core member Feng Zhang, Professor,“said SHERLOCK to provide an inexpensive, easy-to-use and sensitive diagnostic method to detect nucleic acid material—this can mean viruses, tumor DNA, and many other targets. Today, the SHERLOCK improved to provide us with more diagnostic information, and brings us closer to providing a practical application for Deployment Tool.”

the researchers before the show SHERLOCK in a variety of applications in practicality. In this new study, they use SHERLOCK to detect lung cancer patient blood samples in a cell-free tumor DNA, while the detection of synthetic zika virus infection and dengue virus, as well as providing other show.

The Strip on the clear results

the papers Co-first author, Zhang laboratory graduate students Jonathan Gootenberg said,“this new uses SHERLOCK technology to the test strip test results let you observe your target exists in the sample, without the use of instruments. This brings us closer to the field diagnosis.”

these researchers have conceived SHERLOCK has a wide range of uses, thanks to it in nucleic acid target detection aspect of the versatility. Zhang added,“this technique shows it in many health care applications potential, including the diagnosis of patient infection and detection to impart drug resistance or cancer-causing mutation, but it can also be used for industrial and agricultural applications, in these applications, the supply chain in the monitoring step to reduce waste and improve safety.”

basic principles

SHERLOCK the key to success is that one is called Cas13 CRISPR-associated protein, this protein is programmed after binding to a specific RNA fragment. Cas13 of the target can be any gene sequence, including the viral genome, in bacteria that confer antibiotic resistance gene or a cancer-causing mutation. In some cases, once Cas13 positioning and cutting it to the specified target, this enzyme will into Overdrive, so the indiscriminate cutting in the vicinity of the other RNA. In order to develop SHERLOCK, these researchers will this“off-target”activity converted to its advantage, to design��This with DNA and RNA is compatible with the system.

SHERLOCK the diagnostic potential depends on the additional suffering after cutting to generate a signal of the synthesized RNA chain. In Cas13 binding to its initial target, it will cut this synthetic RNA, the release of signal molecules, thereby generating a indicate that it targets the presence or absence of the measurement value.

detection of multiple targets

SHERLOCK platform are now able to apply to test a variety of targets. SHERLOCK was initially at once can only be detected by a nucleic acid sequence, but now a single analysis can simultaneously serve up to 4 different targets to provide a fluorescent signal—this means that you need less samples for the diagnosis. For example, this new SHERLOCK version can be in a single reaction in determining a whether a sample contains a zika virus infection or dengue virus particles, the virus particles will cause the patient to appear similar symptoms. This platform utilizes from different bacterial species of the Cas13 and Cas12a, formerly known as the Cpf1）enzyme to produce these extra signals.

increased sensitivity

SHERLOCK the second iteration also uses an additional CRISPR-associated enzyme that Csm6 to enlarge its detection signal, so that this tool than it’s predecessor having a higher sensitivity. Papers Co-first author, Zhang the Laboratory of the Massachusetts Institute of technology graduate Omar Abudayyeh explained,“the use of the original SHERLOCK, we can detect the 1 L sample in a single molecule, but now we can make this sensitivity increase 100 times. This is for detecting the blood sample in the cell-free tumor DNA or the like of the application is particularly important in these applications, the target molecule concentration may be very low. This new generation of features helps to make SHERLOCK into a more accurate system.” Biological Valley Bioon.com

Published at Fri, 15 Mar 2018 07:00:58 +0000