...overview of these new variants:   Figure 2: Inner mitochondrial membranes imaged with structured...for systemic gene transfer in primate brains Endothelial-specific AAV delivery Recombinant anti-FLAG tag... it with an “assist peptide” that facilitates endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA...10.1038/s41565-023-01419-x. PMID: 37430038. Endothelial-specific AAV delivery By Cythina Arokiaraj (Gradinaru...to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with slightly reduced ...proof-of-concept study, mouse LY6A was delivered to brain endothelial cells in mice lacking LY6A using AAV1-X1 (Chen...X1 and AAV9-X1.1 efficiently transduce brain endothelial cells across diverse mice strains and rats. Figure...

...account when simulating a digest as depicted in Lane 2. Controlling methylation Finally, you can control...prevent them from being degraded by restriction endonucleases. A given prokaryote typically has genes encoding...groups to specific DNA sequences and companion endonucleases that recognizes and cleave the same DNA sequence...

...phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be detected by using a MS2 coat... 296.5569 (2002): 913-916. PubMed PMID: 11988576. 2. Tsien, Roger Y. "The green fluorescent protein." ...requiring anaerobic conditions, proteins that bind endogenous fluorescent co-factors can be used. Examples ...

...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...Choudhury, Rajarshi, et al. "Engineering RNA endonucleases with customized sequence specificities." Nature...

...Drosophila melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169. Soo...

...double strand break), increasing editing efficiencies 2-3-fold. However, the process of double nicking increases... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...Adamson, B. (2024). Improving prime editing with an endogenous small RNA-binding protein. Nature, 628(8008),...

...available in the adenovirus section below!   Fig. 2 - Viral vector components of AAV Adenovirus Adenoviruses...virion assembly and particle release from late endosomes when AAV is trafficked. Rep and cap mediate AAV...Of note, recombinant adenovirus is missing the endogenous E1 gene, making it replication deficient. This...

...split at 175 Arabidopsis homolog of the yeast TREX-2 mRNA export complex: components and anchoring nucleoporin...mNG2(11) Improved split fluorescent proteins for endogenous protein labeling. Feng et al. Nat Commun. 2017...tools for faster, easier fluorescent labeling of endogenous proteins in Caenorhabditis elegans. Goudeau et...1-10) Improved split fluorescent proteins for endogenous protein labeling. Feng et al. Nat Commun. 2017...fluorescent proteins for the visualization of endogenous proteins and synapses. Feng S et al. Commun Biol...et al. Improved split fluorescent proteins for endogenous protein labeling. Nat Commun 8, 370 (2017). https...

...mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct...composed of restriction endonucleases and methyltransferases. Restriction endonucleases cleave DNA at or near...from cleaving the DNA. A role for restriction endonucleases in plasmid addition was first described by studies...stable than the other. Here, the restriction endonuclease is more stable than the methyltransferase. When...unmethylated sites in the chromosome. The more stable endonuclease can then cut the chromosome leading to cell ...

...posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA...about 125 nt that specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB...

...teaching and mentoring others (see more in Figure 2). Starting early demonstrates not only your interest...application process so you can learn more! Figure 2: The many ways to prepare for a science policy career...regulatory approval for a new product or a well-endowed philanthropy who can shift the funding landscape...

...PMID: 27151215. PubMed Central PMCID: PMC4938170. 2. Zuris, John A., et al. "Cationic lipid-mediated delivery...purchase the Cas9 protein and a gRNA from a commercial vendor, this can often be expensive. An alternative approach...from ssDNA, which can be generated by commercial vendors such as IDT. These two components are then incubated.... "Receptor-Mediated Delivery of CRISPR-Cas9 Endonuclease for Cell-Type-Specific Gene Editing." Journal...

...including endothelial cell-pericyte, neuron-astrocyte, and cancer-stromal cell pairs.   Figure 2: Schematic...your protein of interest. TIR1 interacts with endogenous Skp1 and Cul3 proteins to form a SCF E3 ubiquitin...

...the world’s population, we account for an estimated 2% of global plastic generation. Less than 9% of plastics...recycling practices below. Reuse lab cooler boxes Some vendors take back their expanded polystyrene (a.k.a. Styrofoam...

...developed fluorescent-protein based DA sensors (Figure 2). The dLight1 dopamine sensor In 2018, Lin Tian’s ...to gather the following types of data: Report endogenous DA release from a single dopaminergic fiber in...

...start selecting and testing systemic capsids! Figure 2 contains a decision tree for selecting a capsid, while... highlighted below the table.     Figure 2: This decision tree can help guide your choice on ...transfer to the adult brain. Nature Biotechnology, 34(2), 204–209. https://doi.org/10.1038/nbt.3440 Goertsen...toward endothelial cells in rodent CNS AAV-PHP.V1 shows higher transduction in CNS endothelial cells (...exception is the X1 capsid for targeting brain endothelial cells, which is available in both AAV9 and AAV1...capsids derived from it, cross the BBB through an endothelial cell protein called LY6A (Hordeaux et al., 2019...functionally expressed on the surface of brain endothelial cells in C57BL/6J mice, but not in BALB/cJ mice...

...backbone, which expresses the gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene...backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors then used GMAP assembly...engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand...number of tandemly expressed gRNAs respectively) and 2) the size of the deletion. Smaller deletions generally...that tests enhancer sequences outside of their endogenous chromatin environment and relies on the fact ...because it was difficult to fuse the AID tag to endogenous proteins. As described in their recent Cell Reports...genes of interest to generate mAID fusions to endogenous genes. Cell lines with the appropriate fusions...

... PMID: 9537413. PubMed Central PMCID: PMC3010411. 2. Kohli, Manu, et al. "Facile methods for generating... using AAV to introduce epitope tags into the endogenous alleles of the p53 and PTEN tumor suppressor ...naturally occurring cancer-causing mutations into the endogenous allele of the STAG2 tumor suppressor in human...gene, such as the introduction of heterozygous endogenous epitope tags, or the introduction of dominant...dominant mutations of a gene into the endogenous allele in cultured cells. AAV genome editing combined with ... 3. Kim, Jung-Sik, et al. "Epitope tagging of endogenous genes in diverse human cell lines." Nucleic acids...

...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology,...the complex is docked on the membrane of the endoplasmic reticulum. And graduate students, momentarily... DNA strand, and then activate two different endonuclease active sites. Fortunately, there is never just...

...six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini...Bioinformatics 33.7 (2016): 1099-1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to...Additional libraries are also available from commercial vendors. These libraries usually contain 4-12 gRNAs/gene...