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Title: Cas9-Based Local Enrichment and Genomics Sequence Revision of Megabase-Sized Shark IgNAR Loci
Authors: Dong, H
Zhang, Y
Wang, J
Xiang, H
Lv, T
Wei, L
Yang, S
Liu, X
Ren, B
Zhang, X
Liu, L
Cao, J
Wang, M
Yang, N
Keywords: Animals
CRISPR-Associated Protein 9
CRISPR-Cas Systems
Fish Proteins
Genetic Loci
High-Throughput Nucleotide Sequencing
Receptors, Antigen
Sequence Analysis, DNA
Issue Date: 1-Jan-2022
Publisher: The American Association of Immunologists
Citation: Dong, H, Zhang, Y, Wang, J, Xiang, H, Lv, T, Wei, L, Yang, S, Liu, X, Ren, B, Zhang, X, Liu, L, Cao, J, Wang, M, JIAHAI SHI, Yang, N (2022-01-01). Cas9-Based Local Enrichment and Genomics Sequence Revision of Megabase-Sized Shark IgNAR Loci. Journal of immunology (Baltimore, Md. : 1950) 208 (1) : 181-189. ScholarBank@NUS Repository.
Abstract: The 0.8-Mb Ig new Ag receptor (IgNAR) region of the whitespotted bamboo shark (Chiloscyllium plagiosum) is incompletely assembled in Chr_44 of the reference genome. Here we used Cas9-assisted targeting of chromosome segments (CATCH) to enrich the 2 Mb region of the Chr_44 IgNAR loci and sequenced it by PacBio and next-generation sequencing. A fragment >3.13 Mb was isolated intact from the RBCs of sharks. The target was enriched 245.531-fold, and sequences had up to 94% coverage with a 255× mean depth. Compared with the previously published sequences, 20 holes were filled, with a total length of 3508 bp. In addition, we report five potential germline V alleles of IgNAR1 from six sharks that may belong to two clusters of the IgNAR. Our results provide a new method to research the germline of large Ig gene segments, as well as provide the enhanced bamboo shark IgNAR gene loci with fewer gaps.
Source Title: Journal of immunology (Baltimore, Md. : 1950)
ISSN: 0022-1767
DOI: 10.4049/jimmunol.2100844
Appears in Collections:Staff Publications

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