...To use this tool to detect a BRAF gene edit, simply PCR-amplify the BRAF locus from the edited cells (...product should contain the edited BRAF sequence), and PCR-amplify the BRAF locus from control cells (the ... the 428 bp band indicates edits in the BRAF gene in the BRAF-gRNA transduced cells, and no such edits.../Cas9 gene edit. I wanted to inactivate the gene BRAF (a kinase implicated in several human cancers) in...editing, don’t expect a 100% knockout population. BRAF gRNA delivery  Overview: In practice you would select...chose one of my stable pools. Viruses containing BRAF gRNA (BRDN0000561167 and BRDN0001146266) ( 76479... different regions of your gene. I used existing BRAF gRNA viruses, but you can find a list of validated...

...et al., 2009).  Zebrafish behaviors Several behavioral assays can be done on zebrafish at 5 days post ...a high degree of control over time and space. Zebrafish have become a popular model organism because their...well to studies of neuroscience. The larvae of zebrafish are translucent and allow for noninvasive live...short generation times and high fecundity, make zebrafish ideal for high throughput studies of optogenetics...optogenetics.  Common opsins used in zebrafish Optogenetics functions through opsins which are light sensitive...to vertebrates and are commonly introduced in zebrafish through the UAS/Gal4 system for behavioral studies...by acoustic, visual or tactile stimuli, larval zebrafish react by performing a distinct C-bend to escape...

...development. Why have zebrafish become such a popular model organism? Zebrafish (Figure 1) were initially...biology that arose from zebrafish-based investigations. Genetic resources for zebrafish A common approach ...implemented in zebrafish. Developmental biology insights using lineage tracing in zebrafish To highlight...control this process. Additional zebrafish applications and conclusions Zebrafish are also popular outside of...Friedrich Miescher Lab of the Max Planck Society. Zebrafish (Danio rerio) have been used since the 1930’s ...maintain. Also in contrast to mice, the transparent zebrafish embryo develops outside of the mother, making ...embryogenesis. To illustrate some of the reasons that zebrafish have been embraced as a model organism, I’ll outline...

... requested gRNAs including those targeting EGFP, BRAF, and MAP3K4. The gRNAs targeting EGFP can be used...controls for gene editing or gene activation, while the BRAF and MAP3K4 gRNAs allow you to target these widely...

...make melanoma cells resistant to treatment with a BRAF inhibitor. (The test was the perfect complement ...

... al. "Mouse cutaneous melanoma induced by mutant BRaf arises from expansion and dedifferentiation of mature...stage. Zebrabow, or “zebrafish brainbow,” is a collection of transgenic zebrafish lines that allow for...(Hadjieconomou et al., 2011). Cell tracing in zebrafish with Zebrabow Before the generation of Zebrabow...(Pan et al., 2011), Brainbow was also used in zebrafish by injecting a CMV-Brainbow construct at the embryonic...Albert, et al. "Multicolor Brainbow imaging in zebrafish." Cold Spring Harbor Protocols 2011.1 (2011): ...

...compounds in one experiment. Zebrafish (Danio rerio) Since the 1960s, the zebrafish has gradually become an...with a disease have a homolog in zebrafish (Howe et al., 2013). Zebrafish are small, easily maintained as...easily bred, and produce 50-300 eggs at a time. Zebrafish embryos are also laid and fertilized externally...their genomes or to create transgenic animals. Zebrafish are an ideal model organisms for embryonic development...differentiation of β-cells in the the developing zebrafish pancreas. They first tagged these cells using ...PMCID: PMC2736433. Howe, Kerstin, et al. "The zebrafish reference genome sequence and its relationship...

...cells.   Zebrafish experiments with MG-2I-dL5** were performed with embryos and adult fish. Zebrafish are ...experimental settings: HEK293 cell culture and in live zebrafish. For cell culture experiments, only dL5**-expressing....  The video below shows what happens to the zebrafish’s heartbeat and blood circulation of following ...

..., generate a dedicated transgenic line. Using zebrafish as a model organism, the Parton Lab recently created..., termed BLITZ (Biotin Labeling In Transgenic Zebrafish) (Xiong, et al., 2021).    Fig. 1: Schematic...be co-expressed in essentially any transgenic zebrafish line expressing a GFP-tagged version of a protein... BLITZ system was designed and optimized with zebrafish in mind, it can easily be applied to other systems...

...reporter for editing enrichment (TREE) The Wang and Brafman lab developed a transient reporter for editing ...specific loci of interest in the genome. The Wang and Brafman lab showed that GFP positive cells isolated with...editing efficiency. TLDR: TREE from the Wang and Brafman lab uses an engineered BFP variant that converts...SJ, Brookhouser N, Schwarz G, Nguyen T, Wang X, Brafman DA (2019) A transient reporter for editing enrichment...

...and lineage tracing in other systems, such as zebrafish and Drosophila. Why Brainbow? Brainbow highlights... study model organisms such as Drosophila and zebrafish. Brainbow in Drosophila has aided in the mapping...motor neurons and the neuromuscular junction. In zebrafish, the method has become very useful in lineage ...labeling for cell tracing and lineage analysis in zebrafish. Pan YA, Freundlich T, Weissman TA, Schoppik D...

...currently shared amongst the zebrafish community. Don E, et al. Zebrafish. 2017. PubMed PMID: 27631880...allow scientists to quickly and easily generate zebrafish transgenic lines to study neurobiology and neuorodgenerative...in ribonucleoprotein particles (RNPs) to both zebrafish embryos and the mouse inner ear. These exciting...

...roles of three Rab proteins during a key stage of zebrafish development (Aljiboury et al., 2023). Rab family...be difficult to unravel their specific roles. Zebrafish are an excellent model for optogenetic studies...Rab35 coordinate lumen and cilia formation during zebrafish left-right organizer development. PLoS Genetics...

...systems, including E. coli, plants, Drosophila, and zebrafish!   Why use multiplexed gRNAs? By expressing multiple...system, check out this blog post.  Multiplexing in zebrafish Wenbiao Chen Lab Golden Gate Assembly Multiplex...These plasmids allow expression of 2-5 gRNAs in zebrafish. Custom destination vectors are used depending...

...targeting the nervous system, and vectors for zebrafish engineering. You can find additional hot plasmids...Gateway-Compatible Toolbox for quickly generating zebrafish transgenic lines to study neurodegenerative diseases...

...experiments The Li lab used GRABDA in drosophila, zebrafish, and mouse brain slices and freely moving mice...threatening looming stimulus in vivo in transgenic zebrafish Report DA release in live mice over the course...

...used FlipGFP to study apoptosis in HeLa cells, zebrafish embryos, and Drosophila. They treated HeLa cells..., a time consistent with previous methods. In zebrafish, the Shu Lab used FlipGFP to track the spatiotemporal...

...developed the LipoGlo system in zebrafish (Thierer et al, 2019). Zebrafish are a great model for high throughput...

...CreLite for cell tracing using the transgenic zebrafish model, ubi:zebrabow. In this system, cells express...found in both deep and superficial tissues of zebrafish studied, such as the brain, eye, muscle and epithelial...

...other species such as non-human primates [7, 8], zebrafish [9], and c. elegans [10]. If used properly, optogenetics...Wyart. "Optogenetics: a new enlightenment age for zebrafish neurobiology." Developmental neurobiology 72.3...