Rapid CRISPR/Cas9-Mediated Cloning of Full-Length Epstein-Barr Virus Genomes from Latently Infected Cells.
作者： Misako Yajima , Kazufumi Ikuta , Teru Kanda
DOI: 10.3390/V10040171
关键词: Bacterial artificial chromosome 、 Clone (cell biology) 、 DNA sequencing 、 Genome 、 CRISPR 、 Cloning 、 Computational biology 、 Biology 、 Whole genome sequencing 、 Virus
摘要: Herpesviruses have relatively large DNA genomes of more than 150 kb that are difficult to clone and sequence. Bacterial artificial chromosome (BAC) cloning herpesvirus is a powerful technique greatly facilitates whole viral genome sequencing as well functional characterization reconstituted viruses. We describe recently invented technologies for rapid BAC using CRISPR/Cas9-mediated homology-directed repair. focus on recent techniques Epstein-Barr virus (EBV) discuss the possible advantages strategy comparatively with precedent EBV-BAC strategies. also design decisions this technology pitfalls points be improved in future. The obtained clones subjected long-read analysis determine complete EBV sequence including repetitive regions. Rapid determination various strains will contribute understanding their global geographical distribution. This can used disease-associated test hypothesis they special features distinguish them from infect asymptomatically.
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