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..., repressing RecA and overexpressing the NHEJ machinery improved NHEJ accuracy in M. smegmatis (Yan et...also be engaged by variants of the canonical Cas9 approach. A pair of CRISPR guides that flank regions of... break; when two such breaks are introduced near each other, in opposite strands. This “double nickase...

...widely utilized across many different cell types and each one requires different elements for vector propagation...control not only the number of plasmids found in each cell, but also whether the plasmid gets integrated...non-physiological levels. Scientists have tried varied approaches to combat these issues. One method to reduce ...

...identical heavy chains and two identical light chains) each with a constant region and a variable region. The...regions, both through genetic and proteolytic approaches. This comes in handy when, for example, you want...product is a mini-Y with both Fab regions still attached to one another. Size: ~110 kDa. Fab' - These fragments...

...rates for AAVs are generally high enough for this approach to work, although it’s less effective in vivo ... of its biggest advantages. Specificity can be achieved using specialized promoters, Flp or Cre dependent...expression AAVs have to infect cells, hijack host cell machinery, and synthesize a second strand of DNA before ...

...introduced gene or over the native locus? Let’s explore each of the options together. Turn it off Turn it off...inducible expression (more on this below) will both achieve temporary repression of protein expression. Addgene...flavors of activators which can be used with this approach - read more about them on our CRISPRa blog! ...

...spatiotemporal resolution than pharmacological or genetic approaches, but only a few such methods exist and they only... used to turn “on” and “off” MEK1 activity by attaching two dimerizing photoswitchable Dronpa (dron= a...at sites homologous to the locations they were attached to in MEK. The photoswitchable versions of MEK1...

...biochemistry department at the University of Massachusetts Medical School. We are working with postdocs...these salary fluctuations, we are continuing to reach out to institutions with a statement indicating ...to hold. There are institutions in Boston - Massachusetts General Hospital, Mass Eye and Ear Infirmary...

...several methodologies currently available for approaching it. These include the traditional restriction...cloning technologies, as well as several others. Each method is unique and relies on specific components...   Figure 2: Image from http://teachline.ls.huji.ac.il/72682/tutorials/gateway/introduction.html...

...protein turnover in situ in neurons. The classic approach for studying protein turnover is radioactive pulse-chase...immunocytochemistry is perhaps the most common approach. With a good and specific antibody and the proper...the proteins of interest are in close proximity to each other can the DNA strands take part in rolling circle...

...you can easily distinguish each different type of antibody and therefore each target protein. This can ...fluorophore-conjugated secondary antibodies specific to each isotype or species to easily discern the proteins...

...little extra and transfer 100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see .... We then select forward and reverse primers for each sample with the help of DNA-editing software which...glycerol plate on dry ice to the sequencing provider. Each sequencing service will have different requirements...

...routine. Set a goal to read a certain number of papers each week to keep up with the research in your area. ...literature in your field of interest. Set aside some time each day or week to at least scan the titles of what’...alerts from journals you read often. The contents of each issue will be delivered to your inbox as soon as...

...to probe the blot again, almost like new. This approach preserves your sample and saves you from having...your sample. Pro tip! Because some protein is lost each time you strip a blot, you should not compare the...cycles! Just remember that a bit of sample is lost in each round of stripping, so it pays to plan ahead.   ...

...thoroughly decontaminated after use, typically with bleach (plastic) or ethanol (other surfaces). For AAVs...and isopropyl alcohol, 0.5% peracetic acid, 10% bleach, or 1% iodine are useful. Some organizations or...policies – basically, you’ll need to get trained in each new place (and pay attention to the trainings!),...

...it is being inserted: one thousand base pairs on each side, in fact. Later, when we transfect the cells...also find the certificate of analysis provided with each cell line. The editing strategy and design described...

... editing in primary human cells  Cell therapy approaches can use base editors to correct mutations in ...specificity, for example for potential therapeutic approaches, the authors recommend mRNA delivery, the V106W... Susanna Bachle...

... Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells. To do...type specific expression in the retina.  Four approaches for designing synthetic promoters  To build a...permission. In total, this library contained 226 AAVs, each with a different synthetic promoter driving expression...

...can express siRNAs to direct the cellular RNAi machinery to silence target genes. This empty MuLV vector...plasmids (based on the Murine Stem Cell Virus, MSCV) each with a unique 6 nucleotide barcode. This collection... Susanna Bachle...

...running professional development (PD) workshops for teachers; organizing Congressional briefings in Washington...eugenics and research misconduct. Past ethical breaches such as the Tuskegee syphilis experiment, the ...non-technical writing skills by writing for science-outreach organizations or student publications, submitting...

...Relations Board to allow graduate students who work as teaching or research assistants to join unions. The topic...provide a better deal for students in areas such as teaching salaries, healthcare access, and benefits; however...Assistant Professor of Sociology. University of Massachusetts Boston Christine Oslowski, PhD - Biotechnology...