Advanced searches left 3/3

Cas9 Mediated Genome Editing - Crossref

Summarized by Plex Scholar
Last Updated: 26 November 2022

* If you want to update the article please login/register

Targeting double-strand break indel byproducts with secondary guide RNAs improves Cas9 HDR-mediated genome editing efficiencies

Abstract Introduction Through manipulation of the homology-directed repair pathway, programmable double-strand DNA fragments can be used for precision genome editing by two-strand DNA fragments. Indel sequences for a given DSB site are reproducible, and can even be predicted, according to the authors. Multiple gRNAs are needed: A primary gRNA that targets the wild-type genomic sequence; a secondary gRNA that targets the most common indel sequence; and one or two secondary gRNAs that target the most common indel sequence; in effect, the procedure gives a u201csecond chance to edit by HDR-mediated editing.

Source link: https://doi.org/10.1038/s41467-022-29989-9


Induce male sterility by CRISPR/Cas9-mediated mitochondrial genome editing in tobacco

Following the introduction of CRISPR/Cas9 technology, Abstract Genome editing has become more common in animal and plant systems. Direct mitochondrial gene-targeted modifications have been found in plants, a form of male sterility known as cytoplasmic male sterility has been associated with specific mitochondrial genes, but no genes have been found in plants. Our findings indicate that mutation of mtatp9 leads to CMS, and that mitoCRISPR/Cas9 can be used to alter plant mitochondrial genomes.

Source link: https://doi.org/10.21203/rs.3.rs-1977971/v1


Efficient virus-mediated genome editing in cotton using the CRISPR/Cas9 system

Plant virus-mediated sgRNA delivery and expression have significant benefits; sgRNA expression can rapidly expand and accumulate as a result of virus replication and migration, resulting in efficient gene editing results. The VIGE receptor, cotton overexpressing Cas9, was the VIGE receptor used in this study. VIGE with CLCrV-mediated VIGE can not only target and knock out the GhMAPKKK2, GhCLA1, and GhPDS genes subgroup A and D genome sequences but also achieve double mutations of GhCLA1 and GhPDS genes at the same time, but also achieve double mutation of GhCLA1 and GhPDS genes at the same time. In addition, the off-target effect assay showed that the CLCrV-mediated VIGE system not only has high gene editing effectiveness but also high gene editing specificity in cotton, assay. To avoid using tissue culture to produce stable genetic mutants, we further investigated whether the FT-sgRNA route could move sgRNA to cotton apical meristem over long distances.

Source link: https://doi.org/10.3389/fpls.2022.1032799


Construction and Optimization of Herpes Simplex Virus Vectors for Central Nervous System Gene Delivery based on CRISPR/Cas9-mediated Genome Editing

Aims: We want to establish parameters that influence the stability and long-term transgene expression of attenuated HSV-1 vectors and their expression in CNS, as well as optimize the expression cassettes to ensure consistent and consistent expression. The virus latent promoter is widely used to stimulate exogenous gene expression, but the exact parameters affecting the longevity and long-term transgene expression of attenuated HSV-1 vectors have yet to be fully understood. Objective: The investigation was conducted with the intention of using the CRISPR-Cas9 device to produce attenuated HSV-1 vectors and determine the effect of transgene expression and vector safety in transgene expression and vector safety. In the mouse hippocampus gene transduction model, the reporter gene expression and safety profiles of each vector were further investigated. Result: The in vitro cell line analysis revealed that the introduction of a gene expression cassette would interrupt virus gene transcription. According to a mouse hippocampus transducing report, complete expression cassette transference at 2. 0 kb intron may result in more robust and long-time gene expression than other constructs. The foreign gene expression would persist much longer when the gene cassette was located downstream of Exon 1, which suggested a role for LAP2 in sustaining promoter activity during latency, according to previous studies. Moreover, over-transcription of the downstream portion of LAT may lead to continuous activation of the attenuated vectors, which indicates a vital role of LAT in maintaining viral reactivation capability.

Source link: https://doi.org/10.2174/1566523219666210618154326


Expanding the Caenorhabditis elegans auxin-inducible degron system toolkit with internal expression and degradation controls and improved modular constructs for CRISPR/Cas9-mediated genome editing

Abstract The auxin-inducible degron system has emerged as a useful way to conditionally deplete proteins in a variety of organisms and cell types. Here, we introduce a toolkit to enhance the use of the AID device in Caenorhabditis elegans. We have produced a set of sgRNA plasmids with improvements that have been shown to improve editing quality, as well as standardized transgene insert sites on chromosomes I and II. This collection of new TIR1-expressing strains as well as modular, cost-effective cloning vectors serves as a platform for quick assembly of CRISPR/Cas9 repair templates for conditional protein depletion.

Source link: https://doi.org/10.1101/2020.05.12.090217


Targeted mutagenesis of the CYP79D1 gene via CRISPR/Cas9-mediated genome editing results in lower levels of cyanide in cassava

Cassava is the world's most popular food root crop, providing calories to millions of Sub-Saharan African subsistence farmers. The cyanogenic glycoside linamarin present in Cassava leaves and roots in large quantities. Cyide poisoning has been associated with the conversion of the cyanogens to cyanide in the body by prolonged consumption of residual cyanogens. The CRISPR/Cas system, which has been in use for gene function research and crop improvement in recent years, has been the most cost-effective and highly portable genome editing device for gene function research and crop improvement in recent years. Mutations within the MeCYP79D1 locus were discovered by genetic sequencing analysis, revealing that the tested putative transgenic plants contained mutations within the MeCYP79D1 locus, with deletions and substitutions being reported both upstream and downstream of the PAM sequence, respectively. The amounts of linamarin and evolved cyanide present in mecyp79d1 lines' leaves were reduced by seven-fold. Our findings show that CRISPR/Cas9-mediated mutagenesis is as an alternative strategy for plantation of cassava plants with reduced cyanide content.

Source link: https://doi.org/10.3389/fpls.2022.1009860

* 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