...non-dividing cells. Cas13 enzymes also don’t require a PAM sequence at the target locus, making them more flexible...

...their reverse complements were parsed for spCas9 PAM sites (NGG) and then filtered based on two main criteria...

... these systems: St1Cas9 requires a very specific PAM sequence that limits the number of targetable loci...

...site-directed mutagenesis can be used to mutate the PAM sequence (an NGG sequence critical for Cas9 cleavage...

...(3.7) is a SpCas9 mutant that exhibits increased PAM flexibility, and this fusion protein allows for LSD1...

...., 2015 ). Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends after...

...may evolve resistance to CRISPR therapies through PAM or seed sequence mutations. Multiple gRNAs could ...

...stricter sequence requirement at the terminal 3’ PAM site (Fu et al. 2013; Pattanayak et al. 2013). This...

... to test a variety of Cas enzymes with different PAM requirements and editing abilities, such as base ...

...they also optimized the system by modulating the PAM to further tune efficiency of knockdown and developed...

...-inducible system was able to recognize the same PAM and had similar targeting specificity as full-length...

...Within the editing window -50 to +50 bp from the PAM site, CRISPR-X produces point mutations at a rate...

...Neuromodulators like dopamine and norepinephrine have important functions in the brain but have been ...study alpha2-AR. Selectivity Norepinephrine and dopamine are similar molecules, and cross-reactivity is... increased selectivity for norepinephrine over dopamine. All three sensors, however, did display some ...some reaction to high concentrations of dopamine. This cross reactivity will need to be accounted for in ...

...adventure. Many thanks to our guest writer, Pamela J. Hines, PhD! Pamela J. Hines is a Senior Editor at Science...This post was contributed by guest writer Pamela J. Hines PhD, Senior Editor at Science Magazine. Although...French horn, and rhubarb. Follow her on Twitter @Pam_Hines and LinkedIn. Additional Resources on the Addgene...

...measurements of dopamine (113049, 113050) or norepinephrine (123308). The two dopamine sensors offer different...pAAV-Ef1a-fDIO-GCaMP6s Neurotransmitter sensors: Dopamine and norepinephrine Neuroscience research relies... different levels of apparent dopamine affinity, with 113050 offering high apparent affinity, and 113049...

...experiment, ChRger2 was non-invasively delivered to the dopamine producing neurons in the ventral tegmental area...were then allowed to “self-stimulate” their VTA dopaminergic neurons by triggering a burst of light that ...poked their nose into a hole in their cage. Since dopamine is an important neurotransmitter that promotes...behavior, if the channelrhodopsins activate the dopaminergic neurons in the VTA, this should propel the mice...

...anesthesia. In Cre transgenic mice, they targeted the dopamine (DA) neurons of the ventral tegmental area (VTA...for encoding aversive stimuli in the mesolimbic dopamine system." Neuron 101.1 (2019): 133-151. PubMed ...Juarez, Barbara, and Ming-Hu Han. "Diversity of dopaminergic neural circuits in response to drug exposure..., Norman E., et al. "Optogenetic activation of dopamine neurons in the ventral tegmental area induces ...

...fragment with FKBP rapamycin binding domain of mTor (Cas9(N)-FRB-NES) resulting in a rapamycin-inducible Cas9...Addgene: Rapamycin-inducible Cas9 sets (Addgene plasmids 62883 &62884; 62885 & 62886) Rapamycin-inducible...Cas9 for genome editing. Without rapamycin treatment, the Cas9(N)-FRB-NES fragment is actively shuttled...to the nuclear export sequence. Treatment with rapamycin induces Cas9(N)-FRB-NES and Cas9(C)-FKBP-2xNLS...

...18 (end) - Panel Alex Tucker (Ginkgo Bioworks) Pamela Hines (Senior Editor at Science) Edward J. Hall...billions of dollars (Freedman et al 2015). Finally, as Pamela Hines of AAAS points out on the panel, irreproducibility...journalists and scientific experts discussed by Pamela Hines Minisymposium on Reproducibility Booklet ...

...Jessica Polka, a Postdoctoral Research Fellow with Pamela Silver.  Most types of biological motion (whether...Jessica Polka is a Postdoctoral Research Fellow with Pamela Silver in the department of Systems Biology at ...