2023 ACMG Annual Clinical Genetics Meeting Digital Edition: Poster Gallery-Application of CRISPR-Cas9 and next-generation sequencing to resolve highly homologous genes in the human genome

Application of CRISPR-Cas9 and next-generation sequencing to resolve highly homologous genes in the human genome
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The document discusses the use of CRISPR-Cas9 technology in resolving highly homologous genes in the human genome. The introduction explains the challenges faced in sequencing genes with high sequence homology and the limitations of current methods in resolving these genes. The document then describes a proof-of-concept for using the CRISPRclean® technology to deplete homologous sequences in next-generation sequencing libraries. The results section presents data on the effectiveness of CRISPR-Cas9 treatment in depleting homology intervals. It shows the depletion rate of homologous regions in samples NA12878 and NA24631. On average, CRISPR-Cas9 treatment resulted in a nearly 50% reduction in reads across all homology regions. The conclusions highlight the effectiveness of CRISPR-Cas9 depletion in removing genomic fragments homologous to pseudogenes and reducing the number of reads aligning to homologous genomic intervals. The depletion rate generated by CRISPR-Cas9 was consistent across sample types. The document also proposes future work to further increase the percent of fragments and reads removed from homology regions. Figures in the document illustrate the depletion rates of homology intervals, the read alignment to homologous genomics regions, and the process of NGS library construction. Overall, the document demonstrates the potential of CRISPR-Cas9 technology in resolving highly homologous genes in the human genome and highlights the effectiveness of CRISPR-Cas9 depletion in reducing false-positive and false-negative diagnostic errors.

Asset Subtitle

Presenting Author - Madhuri Hegde, PhD, FACMG; Co-Author - Jorune Balciuniene, PhD, FACMG, ErCLG; Co-Author - Fen Guo, PhD, FACMG, FCCMG; Co-Author - Zeqiang Ma, PhD; Co-Author - Keith Brown, PhD; Co-Author - S Choudhary, PhD; Co-Author - A Siddique, PhD; Co-Author - S Ranganathan, PhD; Co-Author - J Armstrong, PhD;

Meta Tag

Chromosome Structure/Function

Clinical Cytogenetics

Comparative Mapping

Cytogenetics

Databases

Education

Exome sequencing

Gene Localization

Gene Mapping

Genetic Testing

Genome sequencing

Genomic Methodologies

Genomic Structure

Methodology

NextGen Sequencing

Pathology

Structure/Function

Co-Author Jorune Balciuniene, PhD, FACMG, ErCLG

Co-Author Fen Guo, PhD, FACMG, FCCMG

Co-Author Zeqiang Ma, PhD

Co-Author Keith Brown, PhD

Co-Author S Choudhary, PhD

Co-Author A Siddique, PhD

Co-Author S Ranganathan, PhD

Co-Author J Armstrong, PhD

Presenting Author Madhuri Hegde, PhD, FACMG

Keywords

CRISPR-Cas9 technology

highly homologous genes

human genome

gene sequencing

CRISPRclean® technology

next-generation sequencing libraries

depletion rate

homologous regions

pseudogenes

NGS library construction