... monitoring sensitized emission in living plant cells." (2013). PubMed PMID: 24194740. PubMed Central ...-molecule FRET for imaging biomolecules in live cells (Sustarsic and Kapanidis 2015), which may lead to...understanding biomolecular structure and dynamics in live cells." Current opinion in structural biology 34 (2015...fluorescent proteins available. Fortunately, several excellent reviews have collated data from various papers... alters the FRET efficiency. Detecting certain cellular changes with an intermolecular probe is often ...Miyawaki et al., 1997), was designed to measure intracellular calcium and published in 1997. Since that time...of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments can probe...

...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...

...Drosophilia Plasmid pRDB_051 (for stable fly cell lines; attL and attR sequences for genome integration...Drosophilia Plasmid pRDB_052 (for stable fly cell lines; Hygromycin resistance gene; for genome integration...rapid assessment of emerging CRISPR technologies. Cell Genomics, 4(3), 100519. https://doi.org/10.1016/...

...or G->A transitions in a target sequence in live cells. This technique uses catalytically inactive CRISPR...and other means of generating mutations in live cells are all discussed further in this informative post...Clearly we have all of our writers to thank for the excellent content they’ve churned out in the last month....

...allowing for direct comparison of Cre+ and Cre- cells. While Cre-lox recombination is a very useful tool...promoter like CAG, or expressed only in a subset of cells under a more specific promoter (e.g. Rho-Cre is ...promoters. Expression of both N and CCre in the same cell type allows for recombination of LoxP flanked DNA...pNrl-Cre Cre-Myc, Expressed in rod photorecetor cells in the retina Nrl Mammalian Cepko 13962 pBS mPrx1... Okano 153207 pAM-AAV-mSncg-Cre Retinal ganglion cell-specific expression of Cre Mouse Sncg AAV Hu 154018...of loxP plasmids, reporter genes indicate which cells have undergone Cre recombination. These plasmids...37402 Cre Stoplight 2.4 Zsgreen is expressed in cells without Cre, mCherry is expressed with Cre Mammalian...

...small Cas protein, is active in E. coli and human cells Two years ago, Jennifer Doudna’s lab discovered ...demonstrating that CasX is active in both E. coli and human cells. Using the structure of CasX at eight different ...

... check viral transgene expression in transduced cells whenever possible. This post will describe our workflow...fragments of DNA from the helper plasmids or the cell genome can be packaged inside the vectors during...earlier, it is common to find DNA from the packaging cell genome, bacterial genomes, cloning vectors, and ...

...overexpress YFG in a Drosophila cell line. Knock downs in Drosophila cell lines are extremely effective....person. Animals are diploid except for their germ cells (sets of genomic material are contributed by mother...

...tag and small molecule. It does work in mammalian cells, mice, and even yeast, though (we have a kit!). ...and complete degradation of proteins in mammalian cells. Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350...Tags Plasmids for Endogenous Gene Tagging in Human Cells Antibodies 101: Epitope Tags  ...uses a small molecule, a tag, and the endogenous cellular degradation machinery to function. The dTAG is...

... localization of that biomolecule inside living cells. FPs classified by the emission color (emission ...proteins and their applications in imaging living cells and tissues”  Physiol. Rev. 90:1103. Wu et. al. ... nuclear localization, and dynamics” Curr. Opin. Cell. Biol. 23:310. .... Therefore, these can be used as “timers” for cellular processes following their activation. The four...Biosensors: Throwing color lights on molecular and cellular processes” Curr. Protein. Pept. Sci. 9(4):338....

... remember what happened when drug X was added to cell line Y, or the phenotype of a mouse with a specific...comparison of the same experiment from different cell lines/model systems ReFigure allows you to make...

...predict that we’d be able to edit RNA in mammalian cells with CRISPR? I certainly didn’t, but now we can ...Improve HDR Efficiency for CRISPR Editing in Human Cells Day 3: Cpf1: Cpf1: A New Tool for CRISPR Genome ...

...example, "while it is possible to grow liver stem cells and liver-like mini-organs or organoids, neither...combine bioengineered scaffolds with patient stem cells to generate compatible functional livers.” Why are... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...

...mismatched translation rates and potential effects on cell growth, even hosts producing alternative tRNAs may...inclusion bodies that will generally segregate with the cell pellet during purification procedures. Even if you...a means to fine-tune gene expression." Molecular cell 59.2 (2015): 149-161. PubMed PMID: 26186290. PubMed...reasons include: Metabolic pressures - it takes cellular resources to produce tRNAs that recognize different...

... . Steps of an IP: 1) Protein extracts from cells or tissues contain a complex mix of proteins seen...common immobilization methods utilizes bacterial cell wall proteins, Protein A and Protein G, which bind...does not. Common negative controls include knockout cell lines or tissues that do not express the protein...

...score (KO-score) which represents the proportion of cells that have either a frameshift or 21+ bp indel. This...Following delivery of CRISPR components into target cells, genomic DNA from both edited and unedited (control... score. KO Score - Represents the proportion of cells that have either a frameshift or 21+ bp indel. This...

...triple colocalization is possible even when staining cells with three IgG1 antibodies. Antibodies were individually...workflow yields similar localization patterns as cells stained for one target (compare nanobodies staining...–rabbit IgG secondary nanobodies. The Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from...

...Control of Gene Expression. Cell 152:1173–1183 . https://doi.org/10.1016/j.cell.2013.02.022 Ronda C, Pedersen... is primarily a means of selection that targets cells in which homologous recombination has not occurred...ensures high editing efficiency; the only non-edited cells to survive have inactivating mutations in the Cas9...one or more genes in E. coli. and in a mammalian cell line (Qi et al., 2013). Two important features of...the bacteria are unable to resolve them, and the cells die.  This could be applied to diseases where the...

...activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the target DNA sequence...activator domain VP64. Using this system in HEK293T cells to induce expression of human IL1RN, the researchers...protein with three red fluorescent tags in U2OS cells. Upon light activation, dCas9 is released within... silencing of transferrin receptor CD71 in HeLa cells.   Comparison of photoactivatable CRISPR strategies...Spatiotemporal Control of CRISPR/Cas9 Function in Cells and Zebrafish using Light‐Activated Guide RNA. Angew...encoded tools, such as microbial opsins, to control cellular activities using light. In 2015, scientists combined...

...about synthetic promoter AAVs for cell-type expression in retinal cells Resources on Addgene.org Find ...