2019
DOI: 10.21859/ijb.2205
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Efficient Production of Biallelic RAG1 Knockout Mouse Embryonic Stem Cell Using CRISPR/Cas9

Abstract: Background Recombination Activating Genes (RAG) mutated embryonic stem cells are (ES) cells which are unable to perform V (D) J recombination. These cells can be used for generation of immunodeficient mouse. Creating biallelic mutations by CRISPR/Cas9 genome editing has emerged as a powerful technique to generate site-specific mutations in different sequences. Objectives The main purposes of this study were to achieve complete knock-out of RAG1 gene by investigating the… Show more

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Cited by 12 publications
(13 citation statements)
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“…These algorithms are typically trained using data obtained from in vitro cleavage assays, which involve using immortalized cell lines ( Störtz and Minary, 2021 ). Immortalized cell lines provide a controlled environment for conducting experiments related to GED ( Mehravar et al, 2019 ), which makes them a valuable resource for training predictive models. It has been observed that these off-target prediction algorithms tend to rely heavily on sequence-based information, which demonstrates a strong connection with the actual cleavage activity of CRISPR/Cas9 or similar gene-editing systems.…”
Section: Ai In Grna Design For Crispr/cas-based Genome Editingmentioning
confidence: 99%
“…These algorithms are typically trained using data obtained from in vitro cleavage assays, which involve using immortalized cell lines ( Störtz and Minary, 2021 ). Immortalized cell lines provide a controlled environment for conducting experiments related to GED ( Mehravar et al, 2019 ), which makes them a valuable resource for training predictive models. It has been observed that these off-target prediction algorithms tend to rely heavily on sequence-based information, which demonstrates a strong connection with the actual cleavage activity of CRISPR/Cas9 or similar gene-editing systems.…”
Section: Ai In Grna Design For Crispr/cas-based Genome Editingmentioning
confidence: 99%
“…Among DNA targeting Cas proteins, Cas9 enzyme guided by single-guide RNA (sgRNA) can specifically bind to target doublestranded DNA (dsDNA), cleaves it due to the nuclease activity, and eventually result in a DNA double-strand breakage. [160,161] The first proof-of-concept study based on CRISPR-Cas9 has been utilized by merging in silicon technology with the NASBA method for primer generation optimal and toehold switches for Zika virus (ZIKV) detection. [161] A microfluidic paper-based analytical device, including freeze-dried reagents for isothermal RNA amplification, in combination with a portable readout device, was used to screen the colorimetric changes upon viral RNA detection.…”
Section: Crispr-cas-powered Biosensorsmentioning
confidence: 99%
“…Biallelic RAG-1 knock-out mES clones (strain 129) had been produced using CRISPR/Cas9 gene editing system and cultured according to the protocols described in our previous work (13). Brie y, mESCs were grown on 0.1%-gelatin-coated culture ask in the absence of feeder cells in mESC culture medium supplemented with R2i and 2% ES-FBS.…”
Section: Gene-targeted Es Cellsmentioning
confidence: 99%
“…Single clones of mESCs were obtained by serial dilution in 96well plates. After clone screening and validation, the pluripotent state of the mutant ES cells was con rmed by Quantitative analysis of pluripotency markers, Nanog, Oct4, and Sox2 genes (13).…”
Section: Gene-targeted Es Cellsmentioning
confidence: 99%
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