...efficiently by (1) improving editing accuracy by preventing re-editing and (2) optimizing “cut-to-mutation... was that these “blocking mutations” mutations prevent CRISPR from re-cutting the target sequence once... mutation incorporation probabilities our data revealed, and found that for generating a homozygous HDR...

...rAAVs, can sound almost too good to be true! In a previous post, we covered systemic capsids, which allow...1, rAAV2/2, and so on, although we often use abbreviated nomenclature (AAV1, AAV2, etc.) to distinguish... known as AAV2-retro) We’ve covered AAVrg in a previous blog post, but to summarize briefly: AAVrg is ...

...cells compared to monomers, although this can be prevented via antibiotic selection (Crozat, 2014). How to... multimerization, and in some cases completely prevent the formation of plasmid multimers (Bedbrook & ...overgrowing the bacterial culture Another way to prevent plasmid multimerization is to avoid growing the...

...gathering data. Former lab members can give open, revealing information about the culture and lab head. While...intimidation, sexual harassment Isolation – including preventing access to opportunities, physical or social isolation...record notes and observations so you have data to review when making a decision. If you don’t see a culture...

...biased and unbiased approaches. As suggested in a review by Tycko et al., 2016, unbiased sequencing and ...High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity." Nature...specificities of CRISPR-Cas9 nucleases." Nature Reviews Genetics17.5 (2016): 300-312. PubMed PMID: 27087594...

...inhibitor of uracil DNA glycosylase (UGI) to dCas9 prevents base excision repair, which would change the U...of inosine from DNA is likely infrequent, thus preventing the induction of base excision repair. In terms...improved base editing capacity, even at sites previously difficult to target. ABE8s can achieve 98-99%...

...that enables high school students to publish peer-reviewed scientific research. You’ll meet some of the folks...mouth and personal networking!) and the manuscript review/submission process at JEI. Chapter 3 - JEI target...

...DeckTheLab and tagging us, @addgene to be judged! In previous years, scientists have decorated their labs with...on the entries, select up to three winners, and reveal the winning entries on Twitter and Instagram! Happy...

...microbiology lab every year for over 20 years. Previously, the lab used three different species for the...continues until all quadrants are used, hopefully revealing single, isolated colonies in the last quadrant...

... expression of specific genes in the brain in reversible fashion. And that’s exactly what the LITEs now...response can be graded with light dose and is fully reversible after light retraction. Finally, because light...

...identify mutations of unknown importance in cancer Previously available methods for investigating and functionally...that that confer proliferation advantages will be revealed in the absence of growth factor. The team assigned...

...honest feedback DePace’s yearly planning meetings revolve around two simple worksheets, each of which is ...sections, the trainee speaks first for each section, preventing the meeting from feeling like an assessment or...

... stage is often much easier said than done. As reviewed in our Plasmids 101 series, proteins are overexpressed...protein expression and purification: a comprehensive review of affinity tags and microbial applications." Biotechnology...

...x-ray machine not on teeth, but on blossoms, thus revealing the startling inner beauty of familiar flowers...such as those collected by Darwin on his voyage. Previous destructive techniques were incompatible with ...

...another is not just to share your riches but to reveal to him his own." -Benjamin Disraeli Mentor, Sponsor...and more. It is instructive (and entertaining) to review this list of bad mentor behaviors. Sadly, it is...

...repression of gene expression in Saccharomyces cerevisiae. The system consists of dCas9, the mammalian ...multiplexed gRNAs in human pluripotent stem cells. Previously, CRISPRi or CRISPRa depended on continued expression...

...assembly from cryo-electron microscopy, the team reveal the mechanism behind its activity including an ...motif, averaging 19 targets per gene. The lab has previously used the library with SpdCas9 to perform pooled...

...As previous blogs have noted, plants are an important foundation for life on Earth. Selective breeding... resistance. Much of plant genetic engineering revolves around Agrobacterium tumifaciens. Agrobacterium...

...transcriptional activation/repression. dCas9 has also previously been adapted to fluorescently label a sequence...calculated fluorescent distances correlating with the previously established physical map. In comparing pairs ...

...Cre-estrogen receptor ligand binding domain-fusion can prevent this toxicity, but with the caveat of partial rather...toxicity. FLP/FRT recombinases (from Saccharomyces cerevisiae) had been used successfully in Drosophila, so...