Cas9-Based Local Enrichment and Genomics Sequence Revision of Megabase-Sized Shark IgNAR Loci.
J Immunol
; 208(1): 181-189, 2022 01 01.
Article
in En
| MEDLINE
| ID: mdl-34880108
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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Sharks
/
Immunoglobulins
/
Receptors, Antigen
/
Fish Proteins
/
Genetic Loci
Limits:
Animals
Language:
En
Journal:
J Immunol
Year:
2022
Document type:
Article