...originally published in Nature, is a “search-and-replace” genome editing technique that mediates targeted... efficient due to its lower dependence on PAM placement. Fewer byproducts and more efficient than homology-directed... formation slightly. Designing the sgRNA with a spacer that only binds the edited strand, as in the PE3b... & Liu, D. R. (2022). Programmable deletion, replacement, integration and inversion of large DNA sequences..., Raguram, A., & Liu, D. R. (2019). Search-and-replace genome editing without double-strand breaks or ...

...Biology and, on occasion, would meet in the same space where BosLab exists today. Back then, it consisted...Kendall square, but we saw the potential in the space and decided to reactivate it. We started out by ...projects at BosLab. Some are entrepreneurs using our space to perform early-stage experiments. For example,... visit our website and follow us on twitter or facebook. If you are local, join us for our next event ...

...both conferences, there was a Q&A section in the interface where you can type in questions for the speaker...posters could be viewed publicly and stored in one place if the participants used the conference hashtag ... says.Depending on the meeting, there may be a “place” for networking within the app. For example, the... an on-site conference, you really have to make space for it,” says Bachle. “On the flip side, you can...

...and novelty in scientific research and in any workspace. Re-design lab meetings with a focus on discussion...making lab meetings more interactive and creating space for discussion at the end of each talk - time for...remotely in science are endless. Let’s adapt and embrace new ways of working, reconnect as human beings ... purposeful and creative collaboration in the workplace. There are many other ways to encourage collaborations...

...example the Phage Assisted Continuous Evolution (PACE) method requires constant turbidity of the culture...its native endonuclease activity degrades the displaced strand.   Figure 1: EvolvR consists of ... of PolI3m. This tag would allow researchers to trace the progeny of a single starting cell. Additionally...generate specific mutations in your plasmids Use REPLACER Mutagenesis to mutate your plasmid Resources ...

... by spaces. File names with more than one word should be named in the camelCaseFormat since spaces between...analyze large data sets (myself included) and we are faced with finding the best system to document these types...the basics. However, these notes take up a lot of space and don’t tell you why you did something or what...

...Personalize all connection requests LinkedIn is a place to build relationships. If you don’t add a personal... talked about (or how good the beer was at that place, for example). This is also a great way to keep ...profile and not updating it. LinkedIn may actually replace the resume some day (I’ve already abandoned my ...have to be a headshot in a suit, but this is not Facebook so no pets or kids. Try looking a bit over your...

...amazing affinity reagents we just didn't have the space to give proper credit to. Thankfully, as Addgene's..., or be able to access an epitope folded into a space that would size-exclude an antibody. In imaging ...eukaryotic cells, allowing for them to be used as intracellular inhibitors.  Aptamers are not proteins at all.../neu and CD16 Exhibit Improved Tumor Lysis When Placed in a Divalent Tumor Antigen Binding Format. Journal...

.... Everything will go wrong Whenever an order is placed on the Addgene website, it sets off a chain reaction... need to be repeated. We also now have a good interface to begin tracking analytics data for lab procedures...at some point, decide early on what to do in a namespace collision. It may even be worth it to have extra... you're solving, but consistency is key. Some namespace collisions you may encounter: vector, sequence...

...repeats separated by an 8-bp long asymmetric core spacer sequence. The asymmetry in the core sequence gives... double-strand DNA break occurs within the core spacer of each loxP site and the two strands are ligated...loxP sites is reversed.   Deletion: If the sites face in the same direction, the sequence between the ...been created, which contain a unique asymmetric spacer "NNNTANNN", where "N" indicates which bases may...

...edit. Figure 2: Cytidine deamination takes place on the free strand of DNA and converts a C to U ..., ABE8s resulted in 1.5-fold more editing at protospacer positions A5-A7 and 3.2-fold more editing at ...Joung’s lab created a dual-deaminase editor called SPACE (synchronous programmable adenine and cytosine editor...mechanism than base editing, has allowed scientists to replace entire sequences of varying lengths. Base editing...and adenine base editors Grünewald et al., 2020 SPACE   Fused cytosine and adenine base editors Walton...

...PubMed search option in early 2020 and eventually replace the existing version of PubMed.  Key features This...references sections of a PubMed entry are great places to look for more articles on your topic of interest...Twitter to promote their work, so it’s also a good place to look for publications. Bonus: papers have to ...automatically update as papers are published, similar to Facebook or Instagram. Key features Setting up a personalized...

..., if these variants are fused to the MLV Gag in place of SpCas9. gRNA A gRNA targeting a DNA sequence ... charge repulsion between virions and the cell surface, and also helps form DNA and retroviral-particle...components. However, this sometimes requires intracellular injections into the cytoplasm or pronucleus ...avoid this pitfall since they are injected into the space surrounding the zygote, where they can then fuse...

...structures, etc., as well as the presence of a Protospacer Adjacent Motif (PAM) sequence. If the gRNA sequence...extent NYN. This increases the flexibility of gRNA placement significantly over SpCas9 and more so than Cas9s...Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science 361,1259-1262(2018). DOI:10.1126/science.aas9129...

...contained all 64 possible NNN PAM sequences adjacent to a protospacer. If ω-dCas9 could recognize a library ...other CRISPR enzymes require a short PAM sequence adjacent to the targeted sequence at the locus of interest... and 3.7 shows that we have only scratched the surface of modifications that can be made to Cas9. It is...

...ultrafugation Overview: Iodixanol gradient ultracentrifugation uses a gradient of different concentrations...during AAV production is overlaid. Following ultracentrifugation, the AAV-containing 40% fraction is collected...day. Refer to the AAV Iodixanol Gradient Ultracentrifugation Protocol for more details. Pro tips Watch...transfer plasmids. It’s easy to contaminate a workspace with plasmid, not that I would know about this...

...quite large (500-600 bp) and may take up precious space in viral transfer vectors with limited packaging...stream of the IRES or 2A element (depending on the placement of the reporter gene). Addgene's collection offers...flanking the multicistronic element(s), or you could replace one of the reporter genes above with your gene ... are designed to have one or more of the genes replaced.  Additionally, 2A peptides could be PCR-cloned...

...gene therapy had fallen - plummeted really -  from grace 2 years prior with the death of Jesse Gelsinger ...these key elements together at the right time and place. But it is the hard work and persistence of a dedicated...early as next year. Twenty years after falling from grace, gene therapy is back, better and safer. It is now...therapy for blindness 2. Gene Therapy emerges from disgrace to be the next big thing, again 3. Acland G. et...

..., the real value here is seeing them all in one place. Can we address problems in scientific publishing...of creating a technology that could eventually replace scientists themselves.  The sections on optogenetics...COVID times,” but the pandemic has certainly exacerbated them and forced scientists to change their priorities...introduction they need to grasp the challenges you face in your career.  Acknowledgements: I would like...

...is then washed, and either eluted from a fixed surface or precipitated, dried, and then rehydrated. There...glass particles in glass milk, gel slices are first placed in a chaotropic salt sodium iodine (NaI) solution...with ddH2O and then ethanol. After regeneration, trace amounts of DNA (3 femtograms/ul) could still be ...cleaned with 1M phosphoric acid and re-used, but trace contamination could interfere with interpreting ...