We narrowed to 803 results for: Cre

...Factors to consider when creating a pooled library amplification protocol. Created with BioRender.com. ...Re-transformation of the SPLINTR Libraries (PDF, 401 KB) Screening Zhang Lab - Multiplexed Overexpression of Regulatory...et al. Nat Protoc. 2017 with modifications Screening pCMV-Rep78/68 Scanning Saturation Mutagenesis...Amplification and Sequencing Protocol (DOCX, 32 KB) Screening Rinehart Lab Iterative Synthetically Phosphorylated...-Cas9 knockout and transcriptional activation screening, a protocol paper by the Zhang lab, and the Broad...More resources on the Addgene blog Genome-Wide Screening Using CRISPR Preparing Pooled Libraries NGS Quality...

...expression? Are you trying to create a single point mutation? Do you want to create a fusion with a reporter...include a homology directed repair (HDR) template to create knock-ins, point mutations, or to add a tag, you...vivo. A common choice for conducting genome-wide screens using CRISPR. AAV transduction Only compatible...Transient expression of CRISPR components, expression decreases as RNA is degraded within the cell. Can be used...Transient expression of CRISPR components, expression decreases as gRNA and Cas9 protein are degraded within the...validated tools.  Special thanks to Joel McDade for creating the Expression Systems table. Marcy Patrick contributed...

...and found that 50 nt spacers increase editing efficiency but also increase off-target editing within the...1090), which decreases the REPAIRv1 construct size from 4,473 bp to 4,152 bp without decreasing editing efficiency..., which deaminate adenosine to inosine in RNA, creating a functional A->G change. They fused ADAR deaminase...to dPspCas13b, but observed low RNA editing. To increase A->G editing, they employed hyperactive ADAR constructs... they hoped to destabilize ADAR-RNA binding to decrease off-targets. Of the tested constructs, ADAR2DD...

...generation oil are moved through small channels to create a water-in-oil emulsion containing approximately...the number of positive droplets as the dilutions increase. The ddPCR software uses the ratio of positive...parameters, here are a few things you can try: Increase the number of cycles. After additional rounds ...amplification, your fluorescence amplitude will increase, leading to greater separation of your positive...droplets. No more than 50 cycles is recommended. Decrease your ramp rate. A rate of 2C/s is recommended ... the droplets, but you can go as low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation...denaturation time to 1 minute has been shown to increase droplet separation (Witte et al., 2016). This is...

...CRISPR screen to disrupt these enhancers. The third and final step is to conduct our CRISPR screen. We start... the marvel of DNA compaction, an enhancer can increase the expression of a gene that is millions of base...stem cells, and conclude with a targeted CRISPR screen that can disrupt these enhancers. A real-world ...that have NANOG binding sites 2. Design a CRISPR screen to target and disrupt the NANOG enhancers 3. CRISPR...you’re interested in designing your own CRISPR screens for enhancers, check out the Appendix. Conclusion...precision medicine. We can apply this knowledge to create targeted disease therapies with minimal side-effects...

...Fluorescent Protein (GFP) has likely existed in creatures such as jellyfish for millions of years; however...physiological processes. Re-engineering GFP to increase its range of color and application Almost as.... Tsien described an S65T point mutation that increased the fluorescence intensity and photostability ...derivatives Mutation Known Function S65T Increased fluorescence, photostability, and a shift of the... the major excitation peak to 488 nm F64L Increased folding efficiency at 37C Y66W Causes the chromophore...Blue-shifts the wavelength (blue derivatives) Y145F Increases quantum yield for BFP Y145A and H148D Stabilizes...to identify particular cell populations in drug screens, to visualize micrometastases in nude mice in cancer...

...Villarreal. By the late 1980s, scientists even created a codon adaptation index (CAI) that is based on... (Sharp and Li, 1987). In 2000 the Ikemura lab created the Codon Usage Tabulated from GenBank database...interest based on an organisms codon usage bias to increase translational efficiency and thus protein expression...across an entire polypeptide. Many researchers have created codon-optimization algorithms/resources (Athey ...study, codon optimization resulted in a 6-9 fold increase in expression of these channels providing scientists...to codon optimize? Although codon optimization increases protein production in certain systems, synonymous...optimization may not always be the optimal strategy for increasing protein production. If you are trying to express...

...s lab performed a series of positive selection screens in bacteria to identify mutants of S. pyogenes ...sequences (Kleinstiver et al. 2015). From these screens, they identified four novel SpCas9 variants with...some cleavage with an NGA PAM. This variant may increase the specificity of genome modifications at DNA...allows for expanded PAM recognition as well as increased specificity and lower off-target activity (Hu ...proteins has and will continue to dramatically increase the number of CRISPR applications. The first non-Cas9...formation of “sticky ends” rather than the blunt ends created by Cas9. Cas12a  displays lower off-target editing...) system fuses Cas13b to RNA deaminase ADAR2 to create a specific RNA editor. Cas13 enzymes are advantageous...

...level of antigen in the sample. This figure was created with BioRender.com  Direct ELISAs A direct ... bind non-specifically to assay components and increase background staining. Direct ELISAs also lack the... antibody. Incorporating a secondary antibody increases the sensitivity of the assay, since multiple secondary... both primary and secondary incubation steps, increasing the workload and time commitment for users. This...antibodies for the capture and detection steps, which increases the specificity of the assay. The assay is also...antibody availability Longer protocolLess specificityIncreased chance of cross-reactivity Sandwich ... High specificityHigh sensitivity Longer protocolIncreased chance of cross-reactivityChallenging to develop...

...they function in nature can inform us on how to create better recombinant vectors. In this blog post, ...provide tips on how we can harness those features to create broad host range recombinant vectors.  A plasmid... host range in plasmids? Not all plasmids were created equally. Only a handful of plasmids have been identified..., from broad host range plasmids can be used to create recombinant vectors that can replicate in different...vinelandii, Brevibacterium methylicum, Caulobacter crescentus, Desulfovibrio vulgaris, Erwinia carotovora, ..., Bordetella spp., Brucella spp., Caulobacter crescentus, Escherichia coli, Gluconacetobacter xylinus,...recombinant vectors can be modified further to increase the range of hosts in which proteins can be expressed...