We narrowed to 587 results for: cel.2;
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TypeBlog Post...much has been done with CRISPR technology in just 2 years and excitement about how much more there is ...alone were worth the trip. Dr. Dana Carroll's excellent keynote address was the first of 19 talks given...
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New and Upcoming Viral Vectors - December 2019
TypeBlog Post...available to the community. We now offer AAV9 for 2 more constructs from Christopher Harvey’s lab. These... results, but also confirm expression in target cells. While the various AAV serotypes can be used to ... -
Plasmids 101: Mammalian Vectors
TypeBlog Post...of translation 2-10 ug/mL G418/Geneticin neo HeLa, NIH3T3, CHO, 293HEK, Jurkat T cells Blocks polypeptide...transfected cells may be considered an entirely new cell line from that of the original parental cells. Positive...cultured mammalian cells. Of course, these mammalian vectors must be compatible with the cell type they are...bacterial cells); however, the techniques and reagents vary. Plasmid transfection into mammalian cells is fairly...expression levels in the cells. Ideally, the chosen reporter is unique to the cell, is expressed from the...components expressed in trans within the cell for effective replication. Cell lines expressing the Epstein–Barr...mammalian cells To achieve stable transfection, there should be a selective pressure to force cells to incorporate... -
Multiplexed Overexpression of Regulatory Factors (MORF) collection
TypeBlog Post...differentiation. Fig. 2: Single cell RNA-seq clustering of cells transduced with the TF pooled...human embryonic stem cells. Transduced stem cells were differentiated and single cell RNA-seq was performed...transduced human embryonic stem cells with the viral TF library, with each cell receiving only 1 TF expression...construct. Cells were cultured following a differentiation protocol before performing single-cell RNA sequencing... 206 cells per TF ORF. From this large sequencing endeavor, a map of possible TF-induced cellular states... clusters of individual cells along with color coded nominated reference cell types. (Adapted from Joung...delivered to each cell, future applications could deliver multiple TFs to each cell, allowing for combinatorial... -
Improving Transduction Efficiency with Exosome AAVs
TypeBlog Post...enrich the microvesicles containing AAVs (Figure 2). Since the development of the exo-AAV isolation protocol...the producer cells and purification protocols often describe a process that begins with cell lysis to release...particles is able to infect more cells (as demonstrated in different cellular environments such as retina ... steps to separate cell culture supernatant containing exo-AVVs from producer cells, and then to enrich...Exosomal vesicles secreted from producer cells also contain cellular macromolecules (e.g. proteins, nucleic...effectively target less accessible cells, such as brain tumor stromal cells in a mouse model of glioblastoma...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector production... -
Antibodies 101: Epitope Tags
TypeBlog Post...is the most charged, with 5 negatively charged and 2 positively charged amino acids. FLAG tag has been .... This tag is not suitable for use in apoptotic cells as Caspase3/7 both cleave after the DVPD sequence...fluorescent marker. The GFP tag is ideal for live cell imaging, FRET, flow cytometry, or immunofluorescence... -
Don’t Wait To Launch Your Health Science Startup
TypeBlog Post...yourself greater chances for success. Build a team of 2 or 3 Don’t go it alone. You’ll find that no matter...would rather fund excellent people with mediocre ideas than mediocre people with excellent ideas. I am lucky... -
FPbase: A new community-editable fluorescent protein database
TypeBlog Post...incorporation of biosensors, protein evolution lineages, 2-photon spectra, more elaborate spectrum analysis tools... There are many online guides – including an excellent series of posts by Joachim Goedhart on the Addgene...Associate at Harvard Medical School. He manages the Cell Biology Microscopy Facility, specializing in advanced... -
Antibodies 101: Monoclonal Antibodies
TypeBlog Post...create a recombinant antibody. Figure 2: Changes to the heavy chain or light chain genes within...isolating the B cells from an immunized animal, those B cells are fused to myeloma cells at a 1:1 ratio...fusion between an antibody-producing B cell and a myeloma cell (National Research Council (US) Committee...immortal cell population that will continue to produce the specific antibody that the original B cell produced... selects against unfused myeloma cells and antibody-producing B cells that cannot replicate, meaning that...monoclonal antibodies are derived from a single B cell clone. One common way of producing monoclonal antibodies...with BioRender.com. The antibody-producing B cells that are used to make hybridomas come from animals... -
Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools
TypeBlog Post...library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express Cas9 and...genome-wide interrogation of the genes required for cellular processes. For a successful screen, it is necessary... on the method used to introduce the gRNAs into cells. Most CRISPR pooled libraries have been created ...the chance of transducing a only a single gRNA per cell. The viral DNA containing the gRNA then integrates...next-generation sequencing of the genomic DNA. However, some cell types are poorly transduced by lentivirus, and thus...CRISPR libraries was not possible. Primary murine T cells cannot be transduced by lentivirus and have been...CRISPR pooled library can be used for screening of cells that are refractory to lentiviral transduction, ...