Invasion by exogenous RNA: cellular defense strategies and implications for RNA inference

Tang, Danxu; Liu, Yan; Wang, Chundi; Li, Lifang; Al-Farraj, Saleh A.; Chen, Xiao; Yan, Ying

doi:10.1007/s42995-023-00209-7

Cited by 4 publications

(1 citation statement)

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“…A similar situation has also been found in insects, where some species are highly susceptible to RNAi while others need large amounts of dsRNA to elicit even a minor knockdown response 61 – 63 . The variability between and within species may be influenced by various factors such as different efficiencies or activities in response to RNAi mechanisms 64 , 65 , limited absorption of dsRNA, or dsRNA degradation in vivo 66 , 67 . Regardless of the underlying reason, it is clear that it is necessary to increase the concentration of dsRNA and providing fresh RNAi bacterial solution in a timely manner to achieve an efficient RNAi silencing in P. caudatum .…”

Section: Discussionmentioning

confidence: 99%

Application of RNA interference and protein localization to investigate housekeeping and developmentally regulated genes in the emerging model protozoan Paramecium caudatum

Gao,

Solberg,

Wang

et al. 2024

Commun Biol

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Unicellular eukaryotes represent tremendous evolutionary diversity. However, the molecular mechanisms underlying this diversity remain largely unexplored, partly due to a limitation of genetic tools to only a few model species. Paramecium caudatum is a well-known unicellular eukaryote with an unexpectedly large germline genome, of which only two percent is retained in the somatic genome following sexual processes, revealing extensive DNA elimination. However, further progress in understanding the molecular mechanisms governing this process is hampered by a lack of suitable genetic tools. Here, we report the successful application of gene knockdown and protein localization methods to interrogate the function of both housekeeping and developmentally regulated genes in P. caudatum. Using these methods, we achieved the expected phenotypes upon RNAi by feeding, and determined the localization of these proteins by microinjection of fusion constructs containing fluorescent protein or antibody tags. Lastly, we used these methods to reveal that P. caudatum PiggyMac, a domesticated piggyBac transposase, is essential for sexual development, and is likely to be an active transposase directly involved in DNA cleavage. The application of these methods lays the groundwork for future studies of gene function in P. caudatum and can be used to answer important biological questions in the future.

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