PT  - JOURNAL ARTICLE
AU  - Bo Zhou
AU  - Steve S. Ho
AU  - Louis C. Leung
AU  - Thomas R. Ward
AU  - Marcus Ho
AU  - Melanie J. Plastini
AU  - Scott C. Vermilyea
AU  - Marina E. Emborg
AU  - Thaddeus G. Golos
AU  - Megan A. Albertelli
AU  - Philippe Mourrain
AU  - Dimitri Perrin
AU  - Karen J. Parker
AU  - Alexander E. Urban
TI  - Haplotype-phased common marmoset embryonic stem cells for genome editing using CRISPR/Cas9
AID  - 10.1101/373886
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 373886
4099  - http://biorxiv.org/content/early/2020/10/21/373886.short
4100  - http://biorxiv.org/content/early/2020/10/21/373886.full
AB  - Due to anatomical and physiological similarities to humans, the common marmoset (Callithrix jacchus) is an ideal organism for the study human diseases. Researchers are currently leveraging genome-editing technologies such as CRISPR/Cas9 to genetically engineer marmosets for the in vivo biomedical modeling of human neuropsychiatric and neurodegenerative diseases. The genome characterization of these cell lines greatly reinforces these transgenic efforts. It also provides the genomic contexts required for the accurate interpretation of functional genomics data. We performed haplotype-resolved whole-genome characterization for marmoset ESC line cj367 from the Wisconsin National Primate Research Center. This is the first haplotype-resolved analysis of a marmoset genome and the first whole-genome characterization of any marmoset ESC line. We identified and phased single-nucleotide variants (SNVs) and Indels across the genome. By leveraging this haplotype information, we then compiled a list of cj367 ESC allele-specific CRISPR targeting sites. Furthermore, we demonstrated successful Cas9 Endonuclease Dead (dCas9) expression and targeted localization in cj367 as well as sustained pluripotency after dCas9 transfection by teratoma assay. Lastly, we show that these ESCs can be directly induced into functional neurons in a rapid, single-step process. Our study provides a valuable set of genomic resources for primate transgenics in this post-genome era.Competing Interest StatementThe authors have declared no competing interest.