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Cas9 - OSTI GOV

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Last Updated: 10 January 2023

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Genome-wide functional screens enable the prediction of high activity CRISPR-Cas9 and -Cas12a guides in Yarrowia lipolytica

Abstract Genome-wide genetic screens have been very successful in determining genotype-phenotype associations and in engineering new phenotypes. Here, we present an experimental-computational approach to sgRNA design that is specific to an organism of choice, in this case the oleaginous yeast Yarrowia lipolytica. The preferred DNA repair technique, non-homologous end-joining, was used to create single guide RNA activity profiles for both SpCas9 and LbCas12a. DeepGuide's report of an organ specific predictor of CRISPR guide activity with retraining can be used to other fungal species, prokaryotes, and other non-conventional organisms.

Source link: https://www.osti.gov/biblio/1904369


Using CRISPR-Cas9 Technology to Eliminate Xyloglucan in Tobacco Cell Walls and Change the Uptake and Translocation of Inorganic Arsenic

We found two Nicotiana tabacum genes that code xyloglucan-specific xylosyltransferases, which we named NtXXT1 and NtXXT2. To see if stopping xyloglucan synthesis would stunt plant growth and plant growth as a result of ageing, we used CRISPR-Cas9 technology to produce ntxxt1, ntxxt2, and ntxxt1/2 mutant tobacco plants. Because no apparent xyloglucan were present in the cell walls of the ntxxt1/2 double mutant's cell walls, we show that NtXXT1 and NtXXT2 are required for xyloglucan biosynthesis. The tobacco double mutant and the related Arabidopsis mutant do not have significant growth defects, but they do have a short root hair phenotype and a slow growth rate. According to Asification and Assignation of As, growing ntxxt mutants in the presence of AsIII or AsV demonstrated that the absence of cell wall xyloglucan enhances As.

Source link: https://www.osti.gov/biblio/1904500


Cas9-derived peptides presented by MHC Class II that elicit proliferation of CD4+ T-cells

Several studies have shown pre-existing immune responses to Cas9, which may have ramifications for clinical development of CRISPR-Cas9-mediated gene therapy. In addition, we can now identify SaCas9 peptides distributed by MHC Class II proteins on dendritic cells using an MHC Associated Peptide Proteomics assay.

Source link: https://www.osti.gov/biblio/1905001


Simple, efficient and open-source CRISPR/Cas9 strategy for multi-site genome editing in Populus tremula × alba

Here, we discuss an effective gene editing tactic for hybrid poplars based on the Golden Gate MoClo cloning program. To mutate the SHORT ROOT gene, two single guide RNAs were created and incorporated into the MoClo Tool Kit level 2 binary vector with the Cas9 expression cassette to determine the system's ability for producing single gene mutants. In addition, we also tested its effectiveness for inserting mutations in two genes simultaneously by expressing one sgRNA targeting a single site of the YUC4 gene and the other sgRNA targeting the PLT1 gene. Transgenic roots for the SHR gene with biallelic mutations for the SHR gene lacked a single cell layer, implying a conserved gene function similar to its homolog, Arabidopsis thaliana Heynh, according to the phenotype. In three and two of the four transgenic roots tested, biallelic mutations for both genes in the yuc4/plt1 and lbd12/lbd4 roots were found in three and two out of three transgenic roots tested.

Source link: https://www.osti.gov/biblio/1904395


CRISPR/Cas9-mediated Targeted Mutagenesis for Functional Genomics Research of Crassulacean Acid Metabolism Plants

The CAM research community is struggling to figure out how the annotated genes in CAM plant genomes plays a huge obstacle. We show that CRISPR/Cas9 is able to induce biallelic indel mutagenesis, revealing previously unknown roles of blue light receptor phototropin 2 in the CAM pathway. Knocking-out of the KfePHOT2 reduced stomatal conductance and CO 2 fixation in the late afternoon and elevated stomatal conductance and CO 2 fixation in the early evening, indicating that blue light signaling plays a key role in the CAM route.

Source link: https://www.osti.gov/biblio/1569393

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions