Noninvasive genotyping of common marmoset (Callithrix jacchus) by fingernail PCR

Abstract

The common marmoset (Callithrix jacchus) is a New World primate that is a useful model for medical studies. In this study, we report a convenient, reliable, and noninvasive procedure to genotype a living common marmoset by using fingernails. This method was used to successfully genotype DNA by restriction fragment length polymorphism (RFLP) PCR without prior purification, by using the KOD FX PCR enzyme kit. Additionally, there is no sample contamination from hematopoietic chimera derived from fused placenta in utero. We compared chimeric levels between various tissues in females with male littermates using quantitative fluorescent (QF)-PCR to prepare a reliable DNA source for genetic analyses, such as genotyping, gene mapping, or genomic sequencing. The chimerism detected appeared to be restricted to lymphatic tissues, such as bone marrow, thymus, spleen, lymph nodes and blood cells. As a result, DNA from fingernails with the quick is the best DNA source for genetic research in living marmosets.

Generation and Breeding of EGFP-Transgenic Marmoset Monkeys: Cell Chimerism and Implications for Disease Modeling

“…Our study rather supports the view that the detected chimerism in marmosets is based on lymphocytic infiltration and not on tissue-specific cell chimerism. This view is also strongly supported by studies using fingernail DNA for genotyping [33,59]. The transgene-chimeric littermates obtained in our study may represent a valuable resource for chimerism studies in marmosets.…”

“…Our data also indicate that cell chimerism between siblings occurs regularly and not only occasionally. Furthermore, our data confirm that the material used for genotyping of (GM) marmosets must be carefully selected to prevent misleading results, e.g., tips of fingernails [33,59] or carefully prepared skin samples.…”

“…In marmosets, blood cell chimerism occurs due to the exchange of hematopoietic stem cells through placental blood vessel anastomoses between siblings [55,56]. Moreover, it is still disputed whether other organs than the hematopoietic system are also chimeric in marmosets [57][58][59]. Therefore, we cultured skin fibroblasts of the F1 animals (except #110 and #111, which were not available for fibroblast analysis) for at least 5 days after a freezing/thawing cycle to deplete the fibroblast cultures of potentially contaminating blood-derived (including chimeric) immune cells.…”

Normal tension glaucoma-like degeneration of the visual system in aged marmosets

“…DNA from marmoset blood cannot be used for genetic analyses as hematopoietic cells are exchanged between siblings in utero , resulting in chimerism 65,66 . Therefore, we extracted DNA from marmoset fingernails 67 . We collected fingernails without the hyponychium.…”

Genetic characterization of a captive marmoset (Callithrix jacchus) colony using genotype‐by‐sequencing

“…A special consideration when interpreting marmoset genomic data is that marmoset littermates are chimeric (Benirschke et al, 1962). Marmoset tissues have a range of chimerism, with hematopoietic-derived tissues showing more extensive chimerism (Ross et al, 2007;Sweeney et al, 2012;Takabayashi & Katoh, 2015).…”