...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...