Efficient Mutagenesis and Genotyping of Maize Inbreds Using Biolistics, Multiplex CRISPR/Cas9 Editing, and Indel-Selective PCR

Themes: Feedstock Production

Keywords: Gene Editing, Genome Engineering, Genotyping

Citation

Rai, M.N., Rhodes, B.H., Jinga, S., Kanchupati, P., Ross, E.H., Carlson, S.R., Moose, S.P. March 28, 2025. “Genome seq of edited H99 callus in maize.” NIH – NCBI.

Overview

Multiplex genome editing of LW1 in ILP1 inbred maize line and Indel-Selective PCR assay design for genotyping.

To identify and quantify any off-target mutations created during the NRT1.1 CRISPR/Cas9 experiment, genomic DNA was extracted from NRT1.1-E1 callus tissue for deep sequencing. Genomic library preparation was conducted using the Nextera DNA Flex Library Prep Kit (Illumina Cat #: 20018704) according to the manufacturers protocol and sequencing was performed on an Illumina NovaSeq6000.

Data

NIH – NCBI: SRX26347042: Genome sequence of edited H99 callus in maize

Download (3.7 KB) includes:

  • Primers
  • Transformation efficiency
  • CRISPR-Cas9 editing outcomes
  • Alignment results

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