CRISPR-Cas9 Gene Editing in Lizards Through Microinjection of Unfertilized Oocytes

Rasys, Ashley M.; Park, Sungdae; Ball, Rebecca; Alcala, Aaron J.; Lauderdale, James D.; Menke, Douglas B.

doi:10.1101/591446

Cited by 21 publications

(23 citation statements)

References 14 publications

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“…This will require the development of new molecular tools, but similar experiments have tested gene function in a new species by leveraging a closely related model organism ( Kamberov et al. 2013 ) and CRISPR transgenic tools are actively being developed for nontraditional model vertebrates ( Rasys et al. 2019 ) making these plausible next steps.…”

Section: Discussionmentioning

confidence: 99%

Molecular Signatures of Placentation and Secretion Uncovered in Poeciliopsis Maternal Follicles

Guernsey

Kruistum

Reznick

et al. 2020

Molecular Biology and Evolution

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Placentation evolved many times independently in vertebrates. Although the core functions of all placentas are similar, we know less about how this similarity extends to the molecular level. Here, we study Poeciliopsis, a unique genus of live-bearing fish that have independently evolved complex placental structures at least three times. The maternal follicle is a key component of these structures. It envelops yolk-rich eggs and is morphologically simple in lecithotrophic species but has elaborate villous structures in matrotrophic species. Through sequencing, the follicle transcriptome of a matrotrophic, Poeciliopsis retropinna, and lecithotrophic, P. turrubarensis, species we found genes known to be critical for placenta function expressed in both species despite their difference in complexity. Additionally, when we compare the transcriptome of different river populations of P. retropinna, known to vary in maternal provisioning, we find differential expression of secretory genes expressed specifically in the top layer of villi cells in the maternal follicle. This provides some of the first evidence that the placental structures of Poeciliopsis function using a secretory mechanism rather than direct contact with maternal circulation. Finally, when we look at the expression of placenta proteins at the maternal–fetal interface of a larger sampling of Poeciliopsis species, we find expression of key maternal and fetal placenta proteins in their cognate tissue types of all species, but follicle expression of prolactin is restricted to only matrotrophic species. Taken together, we suggest that all Poeciliopsis follicles are poised for placenta function but require expression of key genes to form secretory villi.

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Section: Discussionmentioning

confidence: 99%

Molecular Signatures of Placentation and Secretion Uncovered in Poeciliopsis Maternal Follicles

Guernsey

Kruistum

Reznick

et al. 2020

Molecular Biology and Evolution

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“…Ultimately the generation of a well annotated reference genome for veiled chameleons will facilitate long‐term genetic based lineage tracing of neural crest cells throughout their induction, migration and differentiation. Such transgenic manipulation is expected to be possible through CRISPR‐Cas9 technology, as was recently demonstrated to function for the lizard Anolis sagrei …”

Section: Discussionmentioning

confidence: 99%

Filling in the phylogenetic gaps: Induction, migration, and differentiation of neural crest cells in a squamate reptile, the veiled chameleon (Chamaeleo calyptratus)

Díaz

Shylo

Roellig

et al. 2019

Developmental Dynamics

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Neural crest cells comprise a migratory progenitor cell population that differentiate into cell types such as neurons and glia of the peripheral nervous system, pigment cells, hormone secreting cells in glands, and skeletal and connective tissue in the head, thus making important contributions to most tissues and organs throughout the vertebrate body. The evolutionary appearance of neural crest cells is considered synonymous with the origin of vertebrates and their subsequent diversification and radiation. While the comparative biology of neural crest cells has been studied for a century and a half beginning with their discovery by Wilhelm His in 1868, most of our understanding of their development and function has come from a small number of species. Thus, critical gaps exist in our understanding of how neural crest cells mediate evolution and development. This is particularly true with respect to squamate reptiles (lizards, snakes, amphisbaenians), which account for approximately one‐third of all living tetrapods. Here, we present veiled chameleons (Chamaeleo calyptratus) as a model system for studying neural crest cell development in squamates. Chameleons exhibit various morphological specializations associated with an arboreal lifestyle that may have been facilitated through neural crest cells acting as a conduit for evolutionary change.

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“…The recent development of technology for genome manipulation of nonmodel organisms (Rasys et al. ) may enable direct tests of the impact of TE‐mediated cluster elongation on Hox gene expression patterns, chromatin modifications, and other functional genomic aspects of Hox clusters.…”

Section: Discussionmentioning

confidence: 99%

Evolutionary lability inHoxcluster structure and gene expression inAnolislizards

Feiner

2019

Evolution Letters

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Hox genes orchestrate development by patterning the embryonic axis. Vertebrate Hox genes are arranged in four compact clusters, and the spacing between genes is assumed to be crucial for their function. The genomes of squamate reptiles are unusually rich and variable in transposable elements (TEs), and it has been suggested that TE invasion is responsible for the Hox cluster expansion seen in snakes and lizards. Using de novo TE prediction on 17 genomes of lizards and snakes, I show that TE content of Hox clusters are generally 50% lower than genome‐wide TE levels. However, two distantly related lizards of the species‐rich genus Anolis have Hox clusters with a TE content that approaches genomic levels. The age distribution of TEs in Anolis lizards revealed that peaks of TE activity broadly coincide with speciation events. In accordance with theoretical models of Hox cluster regulation, I find that Anolis species with many TEs in their Hox clusters show aberrant Hox gene expression patterns, suggesting a functional link between TE accumulation and embryonic development. These results are consistent with the hypothesis that TEs play a role in developmental processes as well as in evolutionary diversifications.

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CRISPR-Cas9 Gene Editing in Lizards Through Microinjection of Unfertilized Oocytes

Cited by 21 publications

References 14 publications

Molecular Signatures of Placentation and Secretion Uncovered in Poeciliopsis Maternal Follicles

Molecular Signatures of Placentation and Secretion Uncovered in Poeciliopsis Maternal Follicles

Filling in the phylogenetic gaps: Induction, migration, and differentiation of neural crest cells in a squamate reptile, the veiled chameleon (Chamaeleo calyptratus)

Evolutionary lability inHoxcluster structure and gene expression inAnolislizards

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