...photostability of the variants is comparable (lower panel of figure 1). To avoid changes in excitation ...various cyan fluorescent proteins is depicted in figure 1. Of note, the photobleaching rate does not depend ...to be used with the fluorescent protein. In figure 1 we provide an example of such a comparison between...

...separation is maintained by 2 sets of interactions : 1. Interactions between the non-target strand and a ...theoretically decrease off-target cutting. Figure 1: Strand separation and target DNA binding to the Cas9... improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally engineered ...M694A/Q695A/H698A). This new variant displays comparable on-target activity and similar or decreased off-target...

...but also others including pseudorabies virus type 1, vesicular stomatitis virus (VSV; Beier et al., 2011...Skeletal Muscle AAV1, AAV6, AAV7, AAV8, AAV9 Table 1: Common AAV Serotypes and their target tissues (adapted... serotypes capable of entering neurons (see table 1)(Choi et al., 2005; Taymans et al., 2007; Howard et...efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One 8:e76310...Wolfe, J.H. (2014). Adeno-associated virus serotypes 1, 8 and 9 share conserved mechanisms for anterograde...,9 in rat neurons and glia in vitro. Virology 372(1), 24–34. PubMed PMID: 18035387. PubMed Central PMCID...transduction efficiency of recombinant AAV serotypes 1, 2, 5 and 8 in the rat nigrostriatal system. J Neurochem...

...require large amounts of high quality DNA. Figure 1 illustrates the three grades of DNA you may need based...their relative quality and quantity.   Figure 1: Plasmid preps fall into different categories based...specific volume ranges of culture as their input (Table 1). When you feel like you need just a bit higher DNA...kit for example. These kits are designed to handle ~1–5 mL of liquid culture. Using a higher volume can ...beads and a suitable magnet as you scale up. Table 1: Standard Prep Kit Capacities* Prep size Amount...Amount of culture Approximate DNA yield** Miniprep 1–5 mL 5–50 µg Midiprep 25–100 mL 20–200 µg Maxiprep...bacteria, a few manufacturers (such as Zymo and TakaraBio) offer yeast-plasmid specific kits.  Medium/High-throughput...

...nucleotides are represented equally, that’s only a 1/16 chance of having a PAM at a given site. While it... NYN/NRN NYN is less effective than NRN Table 1 - Summary of PAM-flexible Cas enzymes Are we truly...editable with CRISPR technology.     Fig. 1: SpCas9 has sparse PAM coverage in the human genome...PAMs and is capable of editing at frequencies comparable to SpCas9. SpRY can also edit NYN PAMs, but with...

...effective recovery of knock-ins, but did result in 1-2 kilobases of additional sequence being introduced...facilitating recovery of edited animals. Figure 1: In the background are 504 restriction digests of ...selection for repair of a temperature-sensitive pha-1 mutation significantly enriches for knock-in of 2x... 3xFLAG epitopes into other, non-linked loci; pha-1(e2123) mutant worms are perfectly viable at 15 ºC,... repair pathway. Reagents required to perform pha-1 co-conversion are available through Addgene. In 2019...brilliantly and irreverently illustrated in the parable of the Geneticist vs the Biochemist, we geneticists...

...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion...and spectral characteristics of the moxFPs are comparable to the non-optimized parental proteins. Our recent...

...restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction enzymes which...The resulting gel image includes a 1kb ladder (lane 1) that has bands ranging from about 500bp to 10kb, ...will allow you to approximate the DNA mass of comparably intense samples of similar size. Run the gel ...

...Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018...and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed Central...Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939-6371-3_3. PMID: 27730575; PMCID: PMC5270762. ... and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed PMID: 6337371. PubMed Central...Tet-Off systems: second generation. https://www.takarabio.com/products/gene-function/tet-inducible-expression-systems...

...another Arabidoposis protein that heterodimerizes with CIBN upon blue light exposure (see Fig 1*). When...fused dCas9 to a truncated CIB1 protein from Arabidopsis (CIBN), which they call the LADL Anchor. Using...

...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9...matured considerably faster, with expression levels comparable to GFP during early embryogenesis (Figure 3)....

... Zebrafish) (Xiong, et al., 2021).    Fig. 1: Schematic showing the workflow for the BLITZ system...Biol. 2022. https://doi.org/10.1038/s41556-021-00836-1 Visual barcodes for live cell clonal multiplexing...SSAP system reports an efficiency of up to 20%, comparable to commonly used CRISPR/Cas9 methods for large...

...all the colors of the CRISPRainbow. Figure 1: CRISPRainbow was used to track the localization of...is available as a kit from Addgene.   References 1. Chen, Baohui, et al. “Dynamic Imaging of Genomic ... labeling efficiency was similar to that of a comparable FISH method. Importantly, they did not observe...

...mammalian expression from human elongation factor 1 alpha Constitutive  Tends to give consistent expression...gene. araBAD General expression Promoter of the arabinose metabolic operon Inducible by arabinose and repressed...

...native peptide bond at the split site. Figure 1: The base editor is split into two plasmids and co-transfected... was also between 1013 - 1014 vg/kg, a dosage comparable to those currently used in human gene therapy...phytochrome-like fusion proteins from the plant Arabidopsis thaliana. In the presence of red light (650 nm...

...editors (ABE) mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make these base changes...dCas to the genomic DNA of interest  Figure 1. When ABEs are in action they bind DNA so that the...found that the smaller version, ABE8.x-m, showed comparable editing behavior to ABE8.x-d. Overall, ABE8s ...

...assumes your data follows. Fig. 1: Examples of comparable groups with and without equal levels of... closely data points fall to each other (see Fig. 1). Alpha values – numerical risk allowance in a ...values from smallest to largest and ranking them (1 being the smallest value). It calculates the cut off...

...depths (1 mm), important for certain applications such as in vivo tissue imaging.   Figure 1: Ace-mScarlet...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and is significantly...

...your finished constructs at Addgene!). References 1. Li, Chaokun, et al. "FastCloning: a highly simplified...actually doing the experiments are also generally comparable so the best way to compare costs is to look at...least 50 cloning reactions. The ligase cost is comparable to that of the restriction enzymes and overall...

...regulation of endogenous RNA. J Biol Eng. 2014 Mar 1;8(1):7. doi: 10.1186/1754-1611-8-7. PubMed.  Related Resources...in the engineering of complex gene networks. A comparable system for targeting specific sequences of RNA...