The team has used VitelloTag successfully in two distantly related species that are important for developmental biology: the sea star (Patiria miniata) and the acorn worm (Saccoglossus kowalevskii). And since their paper went online, “we’ve been getting a lot of interest from people in different institutions, who want to try it in their own particular critter,” Swartz said.
Vitellogenin is highly conserved across animal species, so their tool may work “as is” with many other organisms. “But the nice thing is, we’ve developed a pipeline where we can make customized versions of VitelloTag that may work in a species, if our first iteration doesn’t,” Swartz said.
Microinjection will still be the method of choice for delivering CRISPR-Cas9 in many organisms. Penetrance (the percent of cells that successfully take up the CRISPR cargo) can be as high as 90 percent with microinjection, whereas with VitelloTag, the team achieved about 30 percent penetrance in the sea star and acorn worm.
“But if you can rapidly add VitelloTag to a dish of 500 eggs and get 30 percent penetrance, you’re still doing good,” Swartz said. “And for an animal like the acorn worm, where injection is just so hard, VitelloTag has a lot to offer. You are going to get way better numbers than you ever could before.”
Collaborators on this study include first author D. Nathaniel Clarke of M.I.T., Akshay Kane and Margherita Perillo of the MBL, and Christopher J. Lowe of the Hopkins Marine Station, Stanford University.
Citation:
Clarke, D. Nathaniel et al. (2024) VitelloTag: a tool for high-throughput cargo delivery into oocytes. Development, DOI: 10.1242/dev.202857
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The Marine Biological Laboratory (MBL) is dedicated to scientific discovery – exploring fundamental biology, understanding marine biodiversity and the environment, and informing the human condition through research and education. Founded in Woods Hole, Massachusetts in 1888, the MBL is a private, nonprofit institution and an affiliate of the University of Chicago.