...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...Natronobacterium gregoryi, or NgAgo for short. Addgene has already distributed this plasmid all over the world...Publication at Addgene Cas9 and NgAgo head to head NgAgo has a number of unique features compared to Cas9. These...compare CRISPR and NgAgo, but it seems that NgAgo has lower potential for off-target effects. In Gao et...

...2015 has been an exciting year for the Addgene blog. We’ve released over 80 posts on a diverse array ...Scientists and the Plasmids 101 2nd ed, the 3rd edition has now been released), and have started providing recorded...incredibly happy to see that one of our career posts has made it to the top of the list this year. While we...doing just that. Do you like the direction the blog has taken over the past year? Do you think there are ...

...dual-nuclease Cas9-Cas9 chimeras Scot Wolfe’s lab has developed dual-nuclease Cas9-Cas9 chimeras with higher...been many Cas nucleases developed for plants, it has been difficult to compare the performance of different...recently discovered in 2017. Now Dominik Niopek’s lab has adapted anti-CRISPR proteins for optogenetic control...

... disease once limited to small pockets in Africa has spread to urban areas and across the ocean. A nurse... considered unlikely to change – but the illness has been fatal in 70 percent of the most recent cases... in the New York Times and on his blog, where he has offered to answer readers' questions. If you have...

... and software development has caught up with it. The peer review process has been challenged with many...targetable NGG PAM sites whereas the human genome has about 300 million. With such a sparsely targetable...needs of each individual; the  community as a whole has yet to settle on any true kind of value. I’ve dubbed... genome editing tool, the CRISPR/Cas9 technology has been surrounded by a whirlwind of research activity...significant improvement. As a result, the software piñata has been filled with slight variations on a theme without...evaluate off-targeting, but currently no software has been designed for this purpose. This second area ...findings to help everyone else. CRISPR technology has reached into so many different communities that it...

...certain experimental setups. The brightness of NLuc has caught the attention of researchers wanting to perform...proteins can be excited by Nluc emission and the lab has dubbed these fusions LumiFluor Reporters. The LumiFluor...mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness and improved maturation...with Luciferases can overcome these problems but has been hampered by dim signals from the coupled luciferase...ReNL; plasmid #85203) NLuc fusions. The Nagai lab has additionally deposited more than 25 fusions of the...Pairing NanoLuc® luciferase with fluorescent proteins has allowed researchers to make innovative tools not ... Technical Services Scientist, Kyle Hooper, PhD, has also worked with R&D for product development and ...

...foot shock that has become associated with a visual or auditory cue. The hippocampus has long been known...every other advanced technique in neuroscience it has a bit of a steep learning curve and many opportunities...within your own neuroscience sub-discipline that has published using the technique and ask if they would...sophistication of mouse transgenics. Everything that has been described is intended for the mouse; however... behavioral models. Beyond rodents, optogenetics has even been applied to other species such as non-human...

...marketing campaign! MIT Green Labs has a lot of resources about this, and even has a list of biological samples...are stored long term at -70 °C. My Green Lab also has resources and even a freezer challenge that your ...

... backgrounds all year and want a change? Addgene has you covered. Today we’re releasing 11 bold science-themed...fan of Addgene or our very cute mascots!  Blugene has been helping us make Plasmids 101 videos and needed...background.           Maybe your viral research has taken you in the direction of COVID-19. We have a...

...by Neville Sanjana. This fantastic research tool has been requested from the repository nearly 1,000 times...provided by the Zhang lab. Even though lentiCRISPR v2 has only been available for a little over a year and ...and a half, a small sampling of the research it has already been used for includes work probing mTOR signaling...

...need new epitope tags? Short epitope tags like FLAG, HA, and Myc can adopt a number of different conformations...effects of GFP fusions to inntags or traditional FLAG, HA, and Myc tags. Impressively, IT6-GFP did not reduce...observed with IT5 and IT6 fusions. In contrast, Flag and HA tags had especially detrimental effects on cell size...

...avoid selecting for cells in which the construct has randomly integrated into the genome, a negative selection... opposed to screen for cells where recombination has occurred. Neomycin, puromycin, and hygromycin resistance...still functions normally because one of its exons has simply been replaced with the same sequence flanked...flanked by LoxP sites while the resistance cassette has been placed into an intron.    Figure 4: ...second the floxed exon.   After recombination has occurred, you'll first remove the resistance marker...

... in NCBI’s numerous databases. As our repository has grown over the years (we now have over 60,000 plasmids...we analyze as part of our quality control process has steadily grown. On a busy week, we may need to analyze...our quality control process. Consequently our team has refined our use of the BLAST web browser interface...also enjoy seeing how our quality control process has changed with the introduction of next generation ...BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started guide for BLAST, which...

... makes software good? Fortunately, that question has been thoroughly answered by experts in human-computer...specialized controls available when needed. SnapGene has been engineered to be easy and enjoyable to use. ...protein sequence match even when the DNA sequence has changed. Identifying plasmid control elements Coding... with these extensive efforts, such an algorithm has limitations. For instance, it could miss a common...

...Transcriptional Programs with CRISPR RNA Scaffolds." Adam has also created a video to help scientists visualize...various flux imbalances. The conventional approach has typically been to either systematically delete/overexpress...introduce heterologous genes, but the trend of research has shifted in recent years toward tinkering with regulatory.... Since the technique relies on base pairing, it has promise to be host-independent, beyond the oft published...

... favorite! Our mascot who has supported us over the past eight years now has a new friend to join them...

...agent than BME and has a less… distinctive smell. BME smells like rotting eggs but has a longer shelf life... It doesn’t degrade during long-term storage and has both high binding capacity and high protein retention... specialty low-background PVDF membrane, it also has higher background noise than nitrocellulose membranes... other hand, doesn’t require any pre-wetting and has lower background than PVDF. It is a little quicker...quicker to use, with high binding capacity, though it has lower retention after binding and washes. It’s less...and need to be read fairly soon after the reaction has started. Types of chemiluminescence enzymes The two...darkroom and a developer. Film is highly sensitive but has a limited dynamic range, which means it may take ...

...Cas9-adenosine deaminase fusion Caixia Gao’s lab has developed a new plant base editor from an evolved...in diverse bacteria from various environments. It has the advantage of stable genomic integration and is...Cre or Flp recombinases used to excise markers. It has the advantage that CRISPR efficiency can be controlled...

...studied but it is only the tip of the iceberg as it has been predicted that many others are still unknown...enzymes. In contrast to other Cas enzymes, Cas14 has not been found in bacterial genomes but only in the...add-on to a diagnostic toolkit, called DETECTR, which has been created by the authors. This kit uses Cas12 ...

...My son is an engineer and my daughter has a business degree. From the very start of their undergraduate...reduced impact of scientists and science. The pandemic has demonstrated that, more than ever, we need all of...at University of Massachusetts Medical School. It has been supported by a Steering Committee and working...