...sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism...PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair ...Cas protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both...

...they either trace downstream from the soma (neuron cell body) toward the axon (anterograde tracer) or upstream...particles require receptor-mediated entry into the cell. This means that not all neurons and pathways will...chemogenetic vector directly into the region where the cell bodies of the projection neurons reside.   Impact...

...co-delivering different AAV particles into the same cells in vivo? A: This will be highly dependent on vector...particularly one that can target the epithelial cells in the blood-brain barrier?   A: For liver, we recommend...worth investigating for both Q6 & Q7.For epithelial cells of the blood brain barrier we would currently recommend...and to all our participants for their time and excellent questions.    Additional Resources on the Addgene...

... glutamate release from neurons and other brain cells in vivo. Glutamate plays a variety of roles in synaptic...promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin-1 promoter. Good for neuronal...promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP: Synapsin-1 promoter, FLP-dependent ...sensor to the membrane and displays it on the extracellular side. In addition to the original PDGFR TMD,...

...electrocompetent cells in which the phage recombination machinery is induced using L-arabinose. These cells can be...Enzymes Required No No Yes Yes Lab Made Competent Cells Required Yes No No No Comparison of REPLACR-mutagenesis...

... in therapeutic settings. As a tool in mammalian cell systems, silencing is achieved through the delivery...). Origins and mechanisms of miRNAs and siRNAs . Cell, 136 (4), 642–655. PubMed (Link opens in a new window...

...patterning of cells and tissues is fundamental to development. I'm interested in how cells find their way...competitive CV You don't necessarily need a Nature/Cell/Science paper. You do need multiple first/corresponding... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...

...plasmids is associated with the "Barcoding cells using cell-surface programmable DNA-binding domains" ...

...bacteria, so it can be transformed as-is into cells. The cells then get to work fixing up the “damage” and...nicked plasmid together long enough to get it into cells and let them do the repair work. The key is T4 DNA...

...Mutations. Berger AH, Brooks AN, Wu X, et al. Cancer Cell, 2016. PubMed PMID: 27478040. The lung cancer mutant... profiling of human gene and allele function via Cell Painting. Rohban MH, Elife, 2017. PubMed PMID: 28315521... of MAPK1/ERK2 Missense Mutants. Brenan L et al. Cell Reports, 2016. PubMed PMID: 27760319. PubMed Central...poorly classified function and a well established cellular signaling gene, thus providing a starting point...reference cDNA clones mapping to well-annotated cellular pathways). It wasn’t clear whether gene-agnostic...sequence-confirmed series of clones corresponding to known cellular signaling pathways as part of our Genetic Pathway...functional assays for these mutations and map other cellular functions and phenotypes. Since time is of the...

...toolkit in human cells. Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK. Mol Cell. 2014 May 22;54(...tRNAs are among the most abundant molecules in a cell, this system is clearly also robust enough to ensure...advantageous to instead use a preexisting, conserved cellular machinery to process gRNAs. Hijacking tRNA processing...

... how it works. The findings in a recent issue of Cell demonstrate how PGC-1a1, a transcriptional coactivator... in a commentary about the new work published in Cell Metabolism. But, they note, there is plenty more...JR, Erhardt S, Lindskog M, Ruas JL. doi: 10.1016/j.cell.2014.07.051. PMID: 25259918 Muscle over mind. Moon...

...with heterotrimeric G proteins on the intracellular side of the cell membrane. Heterotrimeric G proteins...Peroxidase-Catalyzed Proximity Labeling. Cell 169:338-349.e11 . https://doi.org/10.1016/j.cell.2017.03.028 Roth BL (2016...small molecules to elicit a response in a certain cell type. Chemogenetics is similar to optogenetics but...induce or inhibit intracellular signaling cascades as a response to the extracellular stimuli. Ligand dissociation...to the seven alpha helices that transverse the cellular membrane. The receptor recognizes the appropriate...signaling is initiated when a ligand binds to the extracellular surface of the GPCR. This results in a conformational...

...Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.org/10.1016/j.cell.2015.09.038 Additional resources... also detect low-frequency cancer mutations from cell-free DNA fragments as well as health-related single...similar types of HPV, HPV16 and HPV18, from human cells, at atomolar levels, within one hour. DETECTR thus...of Diverse Class 2 CRISPR-Cas Systems. Molecular Cell 60:385–397 . https://doi.org/10.1016/j.molcel.2015.10.008...

... brain regions. You can partially control which cells are successfully infected by an AAV by altering ...control where your gene of interest is expressed.  A cell-type specific Cre can provide you with that control...

...mixture of two base editor enzymes in mammalian cells. Read the paper in bioRxiv Find Target-ACE   CRISPR...bionanoparticle Transient expression of CRISPR/Cas9 in cells has advantages like reduced off-target activity ...

...multiple plasmids in one cell, it might be that you can't stably maintain them in the cell. This blog post features...

...pathways that are frequently implicated in cancer cell proliferation, survival, differentiation, and apoptosis...pathway. “The idea is that if you introduce them into cells one at a time, you could identify those that confer...

... GFP, enabling you to select cells with high GFP expression. These cells have the highest levels of Cas9... on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the desired...gRNA, which causes hypopigmentation, thus marking cells that have undergone genomic modification. In this..., et al. “Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system.” Scientific...

...lack uracil, but contain methionine. (This allows cells carrying nonfunctional as well as functional MET... is available at our Investigations in Molecular Cell Biology Website. Many thanks to our guest blogger...right) has taught courses in biochemistry, molecular cell biology and genetics at Boston College. She is particularly...