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bioRxiv · 10.1101/2023.03.17.533097

Creation of Advance Construction of CRISPR/Cas9 via using Multi Round Gateway Cloning

Jameel, M. R.
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Abstract

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 (CRISPR-associated system) is used to edit specific genomic sequences with precision and efficacy. CRISPR/Cas9 system mediated double strand binding site through error-prone non homologous end joining repair in targeted sequences, which the deletion or insertion can be achieved by knock out gene. Creation advance construct of CRISPR/Cas9 of soluble starch synthase II-1, 2 and 3 (SSSII) genes in single destination binary vector through multi round LR reaction of Gateway technology. Gateway cloning system developed the suitable for the site-specific DNA recombination of lambda-bacteriophage and ccdB the cytotoxic protein, which is poisonous to most E. coli strains. A genetic transformation vector designed with appropriate gRNAs, Cas9, and antibiotic resistance was used to create SSSII knockout mutants.

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Jameel, M. R.. 2023-03-18. Creation of Advance Construction of CRISPR/Cas9 via using Multi Round Gateway Cloning. https://doi.org/10.1101/2023.03.17.533097

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