...hard to make out the details. To increase the resolution, the Xu lab created a dCas9-based CAPTURE (CRISPR...enhancers. CAPTURE 2.0’s multiplexing and high resolution allowed the Xu Lab to simultaneously characterize...chromatin's dynamic role in gene regulation The high resolution and multiplexed chromatin ‘photos’ generated ...

...polyethylene glycol (PEG). We add 25 mL of 40% PEG solution to each 100 mL of supernatant (final concentration...need to determine the titer of your viral vector solution. A variety of titering methods have been developed...Boye, S. E., Levites, Y., … Golde, T. E. (2020). Utilizing minimally purified secreted rAAV for rapid and...

...department during the application process. Many institutions recommend that you reach out to potential advisors...you can’t visit. By talking with people at the institution, you can get a better sense of the environment...answer process can also help you to see if the institution is right for you outside of just the program ...

...the use of AdV for in vivo CRISPR-Cas delivery utilized the Non Homologous End Joining (NHEJ) DNA repair..., 2014). An alternative approach to NHEJ is to utilize the Homology Directed Repair (HDR) to knock in ...and very efficient introduction of DNA into therapeutically relevant, non-transformed mammalian cells. ...

...are treated with a drug of interest and gRNA distribution is analyzed in the drug-resistant population... resistance to drugs and can identify future therapeutic targets for diseases causing uncontrolled cell...6 gRNAs per target gene, and maintaining the distribution of each gRNA within the population is key. Loss...

...their problem and piecing together a really great solution that covered all the bases. You may have also ...skills to convey your empathy and your awesome solution to the person over email. Examine your current...Speaking Time management Managing a team Conflict resolution Organization Leadership Problem Solving Creativity...

...They then performed sixteen rounds of directed evolution across the full ADAR2DD to further optimize cytidine...   Addressing the potential use of RESCUE in therapeutic applications, the authors tested various dRanCas13b...of RNA editing in research and its potential therapeutic uses We are also curious if REPAIR and RESCUE...

...unlimited production, reliable expression, and easy distribution as DNA (Trimmer, 2020), you may be interested...The beads are then washed, followed by antibody elution. Whatever method you choose, you may not be able...any temperature shifts. Of course, we are also cautious during the aliquoting and storage process to prevent...

... GCaMP containing plasmids are available for distribution. If your experiment involves delivering a calcium...types of searches including depositing labs and institutions. For example, search for “Melina Fan” and you...Lab published in eLife. Addgene is currently distributing 94 plasmids expressing validated antibodies ...

... challenge. Find the PHP Plasmids Here! Their Solution: Use the CREATE method to engineer two new IV ...stands for Cre Recombinase-based AAV Targeted Evolution. In this technique, a pool of mutant AAV capsid...labeling is to inject a mix of AAV vectors that constitutively express a red, green, or blue (RGB) fluorescent...

... be to fuse an affinity tag to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses...KRRWKKNFIAVSAANRFKKISSSGAL 4 Affinity and Purification Binding and elution steps use very moderate buffer conditions FLAG...cytometry, but rarely used for purification as elution requires low pH poly His HHHHHH  0.8 Affinity...

... variety of fields in order to provide useful solutions to complex challenges. As a cross-disciplinary...citation rates to measure the impact of their contribution. It offers a 5-step process and several examples...Social Networking for Scientists Read about the Evolution of Scientific Impact Data Freedom: The Expansion...

...by Gaudelli et al. resulted in the successful evolution of an adenosine deaminase that can act on DNA ...for surviving colonies and subsequent rounds of evolution and engineering gave rise to a mutant TadA (TadA...in rice. In Genome Engineering: The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional...

...in vivo experiments and in biotechnology and therapeutics (Mauro and Chappell 2014).  Ultimately, it is...critical analysis of codon optimization in human therapeutics. Trends in Molecular Medicine 20:604–613  https...ZE, Kimchi-Sarfaty C (2011) Understanding the contribution of synonymous mutations to human disease. Nat...

...The hardest part of any revolutionary discovery, including CRISPR, is turning potential into impact. ...collaborators of Omar and Jonathan and experts in CRISPR evolution, were also working on the project. Using computational...possibilities at the bench and potentially in therapeutic applications. The potential of CRISPR has yet...

...specific purposes as described previously (Table 1, Gutierrez-Guerrero, et al, 2020). Because retrovirus is ...disadvantages: Table 1 (Joglekar et al., 2017)  Table 1 (Gutierrez-Guerrero et al., 2020) Table 1 (Cronin et al.,...–222 . https://doi.org/10.1006/mthe.2000.0116 Gutierrez-Guerrero A, Cosset F-L, Verhoeyen E (2020) Lentiviral...

...Briefly, 293T cells are transduced with serial dilutions of a lentiviral vector, incubated for 72 hours...line was obtained directly from Takara, and is routinely tested for mycoplasma contamination using mycoplasma...Cells transduced with our lentiviral vectors are routinely checked for signs of microbial contamination. ...

... Variation in ori structure allows plasmids to utilize diverse host replication machinery. For instance...Paracoccous denitrificans, Pseudomonas fluorescens, P. putida, Rhizobium meliloti, R. leguminosarum by. viciae... P. C., & Wong, H. L. (2023). Development of constitutive and IPTG-inducible integron promoter-based expression...

...; multiple codons encode single amino acids. Evolutionary constraints have molded which codons are used...express well in any organism. So what are the evolutionary constraints that lead to these preferences and...6. Pechmann, Sebastian, and Judith Frydman. "Evolutionary conservation of codon optimality reveals hidden...

...resistance marker from the vector (abR, Figure 1). The resolution step (“double crossover”) which leads to the ... program) (Latynski and Valentovich 2014). We routinely clone the purified amplicons upstream and downstream... in 5 days. Counter-selection with sacB has revolutionized allelic exchange. Even now, only half our lab...