...CRISPR plasmids that create double strand breaks in DNA....CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...

...gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes...nickase plasmids for the generation of single-strand breaks. CRISPR...proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous...

...When it comes to breast cancer, Hoshinho and colleagues found that after injecting breast cancer cells into... Xing et al., 2010) revealed precisely that for breast cancer. What are tumor microenvironments made ...proliferation. However, the lack of caveolin 1 in breast cancer also induces changes of stromal cells in...characterization of the tumor microenvironment in breast cancer." Cancer cell 6.1 (2004): 17-32. PubMed ...Myoepithelial cells: autocrine and paracrine suppressors of breast cancer progression." Journal of mammary gland biology...Diana, and Fernando Schmitt. "Microenvironment in breast tumorigenesis: Friend or foe?." Histology and histopathology...tumor recurrence and poor clinical outcome in human breast cancers." The American journal of pathology 174.6...

...mice can be included in breeding strategies identical to those used for breeding heterozygous mice. In ...all of the offspring in the F1 generation, your breeding scheme should start with parental mice that are...detect your targeting vector or gene of interest. 2. Breed your parental mice to generate F1 Once you’ve performed...locus (e.g. Cre) and heterozygous at the other. 3. Breed your F1 generation Now, your goal is to generate...as progeny from future generations. 4. Continue breeding your colony You’ll want to generate enough mice...controls in your experiment - use mice that were bred in your colony, as opposed to ordering control mice...harbor similar epigenetic modifications. Special breeding cases Whether your mouse colony consists of genetically...

...yet? If not, just remember that the next holiday break is just a few weeks away. Is your favorite science...

... 30125270. Changing the way we detect viral outbreaks Article contributed by Izzy Mueller  Listen to...highly contagious, it is essential to contain outbreaks when they surface. But detection of the virus ... podcast segment The human proteome contains a breadth of modular domains that facilitate protein-protein...

... DMEM complete + 10 µg/mL polybrene by diluting 20 µL of 10 mg/mL polybrene into 20 mL media. Thaw the...with DMEM complete + 10 µg/mL polybrene, the final concentration of polybrene in each well should be 10 µg...glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 ...of the lentivirus in DMEM complete + 10 µg/mL polybrene. Note, this is just a sample of possible dilutions...Lentivirus (μL) Volume of DMEM complete + 10 µg/mL polybrene (µL) 0 0 500 1:5 300 200 1:10 150 350 1:50 30 ...solutions at various dilution. Make sure to use the polybrene-containing media to make the cell solution in ...total volume of 7 mL of DMEM complete + 10 µg/mL polybrene. Mix well by pipetting or inverting the tube. ...

...complete containing 10 μg/mL polybrene by diluting 20 μL of 10 mg/mL polybrene into 20 mL of media. If using...glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 ... before transduction. Note that in that case, polybrene should only be added at the time of viral transduction...lentivirus into DMEM complete containing 10 μg/mL polybrene: Dilution Volume of Lentivirus or Previous Dilution...) Volume of DMEM Complete Containing 10 μg/mL Polybrene (μL) Volume of Virus Added to Plate (μL) Volume... 9.45 mL of DMEM complete containing 10 μg/mL polybrene. Mix well by pipetting or inverting. Aliquot 1.35...cells may be more sensitive to lentivirus or to polybrene, in which case this incubation step can be reduced...

...μg/mL polybrene. TIP: Polybrene increases the efficiency of viral infection. However, polybrene is toxic.... Hexadimethrine Bromide (Polybrene) Prepare a 1mg/mL solution of polybrene (Sigma-Aldrich catalog #H9268... Vendor and catalog # Hexadimethrine bromide (polybrene)* Sigma-Aldrich: #H9268 Protamine Sulfate* MP ...your experiment Detailed protocols for preparing polybrene, protamine sulfate, and puromycin are located ... cell lines, substitute protamine sulfate for polybrene. c. Add lentiviral particle solution from step...at 4°C for up to one year. The powder form of polybrene is stable at 4°C for several years. Protamine ...

...EX0276-1 Benzonase 250 U/µl, Millipore #71205-3 Polybrene 10 mg/mL, Millipore, TR-1003-G Molecular Biology...300,000 cells/well in 1350 µL media and 11.1 µg/mL Polybrene into the wells containing the virus and the untransduced...,000 cells in 13.5 mL of media and 11.1 µg/mL Polybrene. Mix the cell suspension well before seeding. ...final volume in the well is 1.5 mL, and the final Polybrene concentration is 10 µg/mL. Mix well and incubate...

...glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene (10mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without...lentivirus into DMEM complete containing 10 μg/mL polybrene. Note, this protocol was developed using low titer...Volume of DMEM complete (μL) Volume of 10mg/mL polybrene (μL) 1:10 150 1348.5 1.5 1:25 60 1438.5 1.5 1:...

...discovered. Browse Plasmids: Single-Strand Break (Nick) , Double-Strand Break (Cut) Increasing Cas9 PAM flexibility...the target DNA. This results in a double-strand break (DSB) within the target DNA, located ∼3–4 nucleotides...small precision edits. NHEJ directly ligates the break ends without the need for a homologous template.... experimentally. Browse Plasmids: Double-Strand Break (Cut) Multiplex Genome Engineering Many CRISPR experiments...Cas3 Rather than initiating DNA cleavage to form a break, Cas3 initiates single-strand DNA degradation. This...running into other proteins bound to DNA, and the break can be repaired by the cell’s endogenous repair ...conversion in RNA or DNA without a DNA double-strand break Cas C RISPR As sociated Protein, includes nucleases...

...gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes...CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...that can alter DNA bases without inducing a DNA break. Cytosine base editors convert C->T (or G->A on ...proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous...

...gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes...CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...that can alter DNA bases without inducing a DNA break. Cytosine base editors convert C->T (or G->A on ...proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous...

...gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes...CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...that can alter DNA bases without inducing a DNA break. Base editors convert C->T (or G->A on the opposite...proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous...

...gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes...CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...that can alter DNA bases without inducing a DNA break. Cytosine base editors convert C->T (or G->A on ...proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous...

...tracrRNA:Cas9 complex unbinds after the double-strand break. CRISPR systems, found in 95% of archaeal and 48%...categories of genome edits. First, the double-strand break (DSB) induced by Cas9 could be repaired through ...repair template with homology arms was supplied, the break could be repaired according to this template, allowing... convert bases without creating a double-strand break. This method displays higher efficiency than HDR-mediated... create permanent genome edits or double-strand breaks. In mammalian cells, Cas13-ADAR2 does not require...

...gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes...CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous...

...minescence‐based T wo‐ Hy brid) : Combines in vivo BRET with ex vivo luminescence-based co-immunoprecipitation...plants. Bioluminescence Resonance Energy Transfer (BRET) CalFlux VNT : An intracellular calcium sensor created...Antares and Enhanced Nano-lanterns) to allow for BRET-based in vivo imaging of multiple cellular processes...ability of a given lncRNA to cis-repress Mauro Calabrese 106253 pLS-mP-Luc Firefly Insertion of 5' promoter...CAG Lentiviral expression of firefly luciferase Brent Stringer 102320 FUW-RLuc-T2A-PuroR Renilla Ubc Lentiviral...

...delivered directly to the brain parenchyma or subretinal space Serotypes The following serotypes were ...enhanced transduction of the retina following subretinal administration and enhanced transduction and ...transduction and lateral spread in the retina following subretinal injection. It was identified by John Chiorini...transduction and lateral spread in the retina following subretinal injection as compared to the parental AAV44.9...