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ABSTRACT: Introduction
Phytic acid (PA) is an important antinutrient agent present in cereal grains which reduces the bioavailability of iron and zinc in human body, causing malnutrition. Inositol pentakisphosphate 2- kinase 1 (IPK1) gene has been reported to be an important gene for PA biosynthesis.Objective
A recent genome editing tool CRISPR/Cas9 has been successfully applied to develop biofortified rice by disrupting IPK1 gene, however, it remained a challenge in wheat. The aim of this study was to biofortify wheat using CRISPR/Cas9.Methods
In this study, we isolated 3 TaIPK1 homeologs in wheat designated as TaIPK1.A, TaIPK1.B and TaIPK1.D and found that the expression abundance of TaIPK1.A was stronger in early stages of grain filling. Using CRISPR/Cas9, we have disrupted TaIPK1.A gene in cv. Borlaug-2016 with two guide RNAs targeting the 1st and 2nd exons.Results
We got several genome-edited lines in the T0 generation at frequencies of 12.7% and 10.8%. Sequencing analysis revealed deletion of 1-23 nucleotides and even an addition of 1 nucleotide in various lines. Analysis of the genome-edited lines revealed a significant decrease in the PA content and an increase in iron and zinc accumulation in grains compared with control plants.Conclusion
Our study demonstrates the potential application of CRISPR/Cas9 technique for the rapid generation of biofortified wheat cultivars.
SUBMITTER: Ibrahim S
PROVIDER: S-EPMC9039650 | biostudies-literature | 2022 Mar
REPOSITORIES: biostudies-literature
Ibrahim Saira S Saleem Bilal B Rehman Nazia N Zafar Syed Adeel SA Naeem Muhammad Kashif MK Khan Muhammad Ramzan MR
Journal of advanced research 20210714
<h4>Introduction</h4>Phytic acid (PA) is an important antinutrient agent present in cereal grains which reduces the bioavailability of iron and zinc in human body, causing malnutrition. <i>Inositol pentakisphosphate 2- kinase 1</i> (<i>IPK1</i>) gene has been reported to be an important gene for PA biosynthesis.<h4>Objective</h4>A recent genome editing tool CRISPR/Cas9 has been successfully applied to develop biofortified rice by disrupting <i>IPK1</i> gene, however, it remained a challenge in w ...[more]