WebJan 26, 2024 · The Cas9 plasmid pCfB2312 was kindly provided by Dr. Irina Borodina (Technical University of Denmark, Denmark) as a part of the “EasyClone-MarkerFree” vector toolkit [ 14 ]. The sgRNA plasmids pCfB9336, pCfB9341, and pCfB9344 were constructed previously [ 15 ]. WebThe publicly available EasyClone-MarkerFree vector suite allows for facile and highly standardized genome engineering, and should be of particular interest to researchers …
Expansion of EasyClone-MarkerFree toolkit for Saccharomyces
WebSep 26, 2024 · The protocol details assembling DNA into integration expression cassettes using the EasyClone method (Jessop-Fabre et al. 2016; Holkenbrink et al. 2024 ), and uses the red phenotype from ADE2 knockout (Ugolini and Bruschi 1996) to demonstrate how to generate and assemble guide RNA vectors for CRISPR-Cas9 genome targeting … WebAug 8, 2024 · Though EasyClone vectors were designed to target 11 well-definedgenomic regions, they were still not suitable for prototrophic strains. Therefore, six different dominant selection markers like nourseothricin, hygromycin were added to EasyClone 2.0 vectors to make them suitable for prototrophic industrial yeasts. Figure 2. cjb therapy center baltimore md
Addgene: EasyClone-MarkerFree: A vector toolkit for marker-less ...
WebJun 8, 2024 · Selection markers are only present on two episomal plasmids, one for Cas9 expression, and another for helper gRNA expression. As no selection markers are … WebThe Expanded EasyClone-MarkerFree toolkit is a new set of 8 validated chromosomal loci for the stable integration of heterologous DNA into baker’s yeast Saccharomyces cerevisiae . This kit will be sent as bacterial glycerol stocks in 96-well plate format. Add to Cart $400 USD + shipping Available to academics and nonprofits only. WebDec 31, 2024 · TL;DR: The EasyClone-MarkerFree vector toolkit as discussed by the authors enables the integration of linearized expression cassettes into defined genomic loci, facilitated by CRISPR/Cas9, which is recruited to the chromosomal location by specific guide RNAs expressed from a set of gRNA helper vectors. dow crash of 2008