...transcription: NP, VP35, VP30, and L (reviewed in 5). The RNA genome is encapsidated by the nucleoprotein, NP. NP...viral mRNAs are translated) by cellular machinery. NP encapsidates the negative sense, single-stranded ...important that all of these plasmids (those containing NP, VP35, VP30, L, and the minigenome) are efficiently...For example, particular ratios of plasmids encoding NP and VP35 allow for efficient polymerase activity,...with appropriate plasmid ratios, we suggest: 500 ng NP, 500 ng VP35, 100 ng VP30, 100 ng L, and 2 μg minigenome...

...sharing landscape as well.     References Andrews NP, Boeckman JX, Manning CF, Nguyen JT, Bechtold H, ...

...10.1038/mtna.2015.32 Maynard LH, Smith O, Tilmans NP, Tham E, Hosseinzadeh S, Tan W, Leenay R, May AP,...

...Addgene.   Find anti-GFP [N86/8R] here!   Andrews NP,  et. al. A toolbox of IgG subclass-switched recombinant...

...recombinant antibody plasmid collection References Andrews NP, Boeckman JX, Manning CF, Nguyen JT, Bechtold H, ...

...yet. References and Resources References Andrews NP, Boeckman JX, Manning CF, Nguyen JT, Bechtold H, ...

...non-denaturing buffer containing non-ionic detergents, such as NP-40 or triton-X, to preserve native protein conformations...

...Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents...

...Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents...

....4.1406. PMID: 4884687; PMCID: PMC225270. Higgins NP, Vologodskii AV. Topological Behavior of Plasmid ...

... are the advantages of Cas9 RNPs? There are many advantages to using RNPs for CRISPR experiments. The...degraded over time, using RNPs may limit the potential for off-target effects. Cas9 RNPs are detectable at high... targeting gRNA, to your cells of interest. Cas9 RNPs are capable of cleaving genomic targets with similar...difficult to transfect, such as primary cells. Using RNPs can also alleviate difficulties with protein expression...via protein degradation pathways. This makes Cas9 RNPs useful for CRISPR applications where limited expression...viral-mediated delivery. You’ve decided to use RNPs, now what? Preparing the Cas9-gRNA ribonucleoprotein...advantage of performing CRISPR experiments using RNPs is the variety of methods that can be used to deliver...

...Thermostability of SpCas9 vs GeoCas9. SpCas9 or GeoCas9 RNPs were incubated in reaction buffer at temperatures...set of experiments tested the ability of GeoCas9 RNPs to be incubated at high temps but still maintain...set of experiments tested the ability of GeoCas9 RNPs to cleave substrate when the reaction was carried...GeoCas9, Harrington et al incubated SpCas9 and GeoCas9 RNPs in diluted human plasma for 8 hours at 37°C. Substrate...longevity of SpCas9 vs GeoCas9. SpCas9 or GeoCas9 RNPs were incubated in human plasma diluted with reaction... may allow for more efficient genome editing via RNPs in vivo. What aspect of GeoCas9 are you most excited...Genome Engineering   Interested in using CRISPR/Cas9 RNPs? Check out this post Check Out Other Blog Posts ...

...particles loaded with Cas9-gRNA ribonucleoproteins (RNPs). According to the authors, they named their tool... carry different types of CRISPR cargo including RNPs for indel formation via NHEJ, precise modification...editing was not detected in cells electroporated with RNPs using the same guide. Nanoblades also had lower ...level of off-target effects, with electroporation of RNPs generally having lower off-target edits compared...

...     Fig. 2: Heatmap indicating loci where SNPs reside on sgRNA target sequences for sgRNAs that...Histograms indicating the frequency (x-axis) which SNPs map to thetargeting sequences of guides across 8...approximately 20% more single nucleotide polymorphisms (SNPs) that map to guides in the Broad’s Genomic Perturbation...lines from European or East Asian ancestries. These SNPs, however, were not accounted for in the CRISPR guides...

...specific Ly6a SNPs on PHP.B binding in vitro. They found that when LY6A contained C57BL/6J SNPs, PHP.B could...Mouse Genome Project. They focused their search on SNPs and small insertions and deletions (indels) most...could bind it, but when LY6A contained BALB/cJ SNPs, binding was reduced. The Deverman group then looked...

... determine whether any sequence variations (e.g. SNPs) exist. Additionally, consider testing multiple ...example, prefer to deliver your CRISPR components as RNPs (ribonucleoproteins) and therefore usually transcribe...

...deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied separately from the viral ...recently showed that by delivering CRISPR components as RNPs and delivering two halves of a repair template spread...

... complexed with sgRNA. Direct comparison of Cas9 RNPs prepared with an in vitro-transcribed (IVT) sgRNA...scaled up these tagging experiments by assembling csRNPs in a 96-well plate format and tested parallel tag...

...specific cell types. Used to identify functional SNPs from loci determined to be important in disease....

...cleaves ssDNA which they have adapted for detecting SNPs. Learn more from this Addgene blog post Read...