Gender Control of Mouse Embryos by Activation of TLR7/8 on X Sperm Via Ligands DsRNA-40 and DsRNA-DR

Overview

Journal Molecules

Publisher MDPI

Specialty Biology

Date 2024 Jan 11

PMID 38202845

Authors

Yunfei Hou

Jingfeng Peng

Linjun Hong

Zhenfang Wu

Enqin Zheng

Zicong Li

Affiliations

Soon will be listed here.

Abstract

Gender control technologies are promising for enhancing the production efficiency of the farm animal industry, and preventing sex-linked hereditary diseases in humans. It has been shown that the X sperm of mammalian animals specifically expresses X-chromosome-derived toll-like receptor 7/8 (TLR7/8), and the activation of TLR7/8 on the X sperm by their agonist, R848, can separate X and Y sperm via the specific inhibition of X sperm motility. The use of R848-preselected sperm for fertilization resulted in sex-ratio-skewed embryos or offspring. In this study, we aimed to investigate whether two other TLR7/8 ligands, double-stranded RNA-40 (dsRNA-40) and double-stranded RNA-DR (dsRNA-DR), are also effective in the separation of mouse X and Y sperm and the subsequent generation of gender-ratio-skewed in vitro fertilization (IVF) embryos. Our results indicated that cholesterol modification significantly enhances the transfection of dsRNA-40 and dsRNA-DR into sperm cells. dsRNA-40 and dsRNA-DR incubation with mouse sperm could separate X and Y sperm by the specific suppression of X sperm motility by decreasing its ATP level and mitochondrial activity. The use of a dsRNA-40- or dsRNA-DR-preselected upper layer of sperm, which predominantly contains high-motility Y sperm, for IVF caused a male-biased sex ratio shift in resulting embryos (with 65.90-74.93% of embryos being male). This study develops a simple new method for the efficient separation of mammalian X and Y sperm, enabling the selective production of male or female progenies.

Citing Articles

Reproductive Tract Mucus May Influence the Sex of Offspring in Cattle: Study in Cows That Have Repeatedly Calved Single-Sex Offspring.

Huang F, Niu P, Wang J, Suo J, Zhang L, Wang J Vet Sci. 2024; 11(11).

PMID: 39591346 PMC: 11598928. DOI: 10.3390/vetsci11110572.

References

Kariko K, Buckstein M, Ni H, Weissman D . Suppression of RNA recognition by Toll-like receptors: the impact of nucleoside modification and the evolutionary origin of RNA. Immunity. 2005; 23(2):165-75. DOI: 10.1016/j.immuni.2005.06.008. View

Hotz C, Treinies M, Mottas I, Rotzer L, Oberson A, Spagnuolo L . Reprogramming of TLR7 signaling enhances antitumor NK and cytotoxic T cell responses. Oncoimmunology. 2016; 5(11):e1232219. PMC: 5139637. DOI: 10.1080/2162402X.2016.1232219. View

Mendez-Acevedo K, Valdes V, Asanov A, Vaca L . A novel family of mammalian transmembrane proteins involved in cholesterol transport. Sci Rep. 2017; 7(1):7450. PMC: 5547113. DOI: 10.1038/s41598-017-07077-z. View

Khramtsova E, Wilson M, Martin J, Winham S, He K, Davis L . Quality control and analytic best practices for testing genetic models of sex differences in large populations. Cell. 2023; 186(10):2044-2061. PMC: 10266536. DOI: 10.1016/j.cell.2023.04.014. View

Elbashir S, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T . Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 2001; 411(6836):494-8. DOI: 10.1038/35078107. View

Obermann H, Lederbogen I, Steele J, Dorna J, Sander L, Engelhardt K . RNA-Cholesterol Nanoparticles Function as Potent Immune Activators TLR7 and TLR8. Front Immunol. 2022; 12:658895. PMC: 8814444. DOI: 10.3389/fimmu.2021.658895. View

Greulich W, Wagner M, Gaidt M, Stafford C, Cheng Y, Linder A . TLR8 Is a Sensor of RNase T2 Degradation Products. Cell. 2019; 179(6):1264-1275.e13. PMC: 7116005. DOI: 10.1016/j.cell.2019.11.001. View

Cran D, Johnson L, Miller N, Cochrane D, Polge C . Production of bovine calves following separation of X- and Y-chromosome bearing sperm and in vitro fertilisation. Vet Rec. 1993; 132(2):40-1. DOI: 10.1136/vr.132.2.40. View

Ohto U, Tanji H, Shimizu T . Structure and function of toll-like receptor 8. Microbes Infect. 2014; 16(4):273-82. DOI: 10.1016/j.micinf.2014.01.007. View

10.

Rath D, Ruiz S, Sieg B . Birth of female piglets following intrauterine insemination of a sow using flow cytometrically sexed boar semen. Vet Rec. 2003; 152(13):400-1. DOI: 10.1136/vr.152.13.400. View

11.

Umehara T, Tsujita N, Shimada M . Activation of Toll-like receptor 7/8 encoded by the X chromosome alters sperm motility and provides a novel simple technology for sexing sperm. PLoS Biol. 2019; 17(8):e3000398. PMC: 6691984. DOI: 10.1371/journal.pbio.3000398. View

12.

Douglas C, Maciulyte V, Zohren J, Snell D, Mahadevaiah S, Ojarikre O . CRISPR-Cas9 effectors facilitate generation of single-sex litters and sex-specific phenotypes. Nat Commun. 2021; 12(1):6926. PMC: 8642469. DOI: 10.1038/s41467-021-27227-2. View

13.

Buchanan B, Seidel Jr G, M McCue P, Schenk J, Herickhoff L, Squires E . Insemination of mares with low numbers of either unsexed or sexed spermatozoa. Theriogenology. 2000; 53(6):1333-44. DOI: 10.1016/S0093-691X(00)00276-4. View

14.

Kagan J, Barton G . Emerging principles governing signal transduction by pattern-recognition receptors. Cold Spring Harb Perspect Biol. 2014; 7(3):a016253. PMC: 4355268. DOI: 10.1101/cshperspect.a016253. View

15.

Guan C, Chernyak N, Dominguez D, Cole L, Zhang B, Mirkin C . RNA-Based Immunostimulatory Liposomal Spherical Nucleic Acids as Potent TLR7/8 Modulators. Small. 2018; 14(49):e1803284. PMC: 6397047. DOI: 10.1002/smll.201803284. View

16.

Heil F, Hemmi H, Hochrein H, Ampenberger F, Kirschning C, Akira S . Species-specific recognition of single-stranded RNA via toll-like receptor 7 and 8. Science. 2004; 303(5663):1526-9. DOI: 10.1126/science.1093620. View

17.

Huang M, Cao X, He Q, Yang H, Chen Y, Zhao J . Alkaline semen diluent combined with R848 for separation and enrichment of dairy goat X-sperm. J Dairy Sci. 2022; 105(12):10020-10032. DOI: 10.3168/jds.2022-22115. View

18.

Khramtsova E, Davis L, Stranger B . The role of sex in the genomics of human complex traits. Nat Rev Genet. 2018; 20(3):173-190. DOI: 10.1038/s41576-018-0083-1. View

19.

Umehara T, Tsujita N, Zhu Z, Ikedo M, Shimada M . A simple sperm-sexing method that activates TLR7/8 on X sperm for the efficient production of sexed mouse or cattle embryos. Nat Protoc. 2020; 15(8):2645-2667. DOI: 10.1038/s41596-020-0348-y. View

20.

Gallas A, Alexander C, Davies M, Puri S, Allen S . Chemistry and formulations for siRNA therapeutics. Chem Soc Rev. 2013; 42(20):7983-97. DOI: 10.1039/c3cs35520a. View