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090			_aB-20870
245	1	0	_aA newly characterized dense granule protein (GRA76)is important for the growth and virulence of Toxoplasma gondii
490	0		_vInternational Journal for Parasitology, 54(2), p.109-121, 2024
520	3		_aPathogenicity of the zoonotic pathogen Toxoplasma gondii largely depends on the secretion of effector proteins into the extracellular milieu and host cell cytosol, including the dense granule proteins (GRAs). The protein-encoding gene TGME49_299780 was previously identified as a contributor to parasite fitness. However, its involvement in parasite growth, virulence and infectivity in vitro and in vivo remains unknown. Here, we comprehensively examined the role of this new protein, termed GRA76, in parasite pathogenicity. Subcellular localization revealed high expression of GRA76 in tachyzoites inside the parasitophorous vacuole (PV). However, its expression was significantly decreased in bradyzoites. A CRISPR-Cas9 approach was used to knock out the gra76 gene in the T. gondii type I RH strain and type II Pru strain. The in vitro plaque assays and intracellular replication showed the involvement of GRA76 in replication of RH and Pru strains. Deletion of the gra76 gene significantly decreased parasite virulence, and reduced the brain cyst burden in mice. Using RNA sequencing, we detected a significant increase in the expression of bradyzoite-associated genes such as BAG1 and LDH2 in the Pru?gra76 strain compared with the wild-type Pru strain. Using an in vitro bradyzoite differentiation assay, we showed that loss of GRA76 significantly increased the propensity for parasites to form bradyzoites. Immunization with Pru?gra76 conferred partial protection against acute and chronic infection in mice. These findings show the important role of GRA76 in the pathogenesis of T. gondii and highlight the potential of Pru?gra76 as a candidate for a live-attenuated vaccine.
650	1	4	_aBRADYZOITE DIFFERENTIATION
650	1	4	_aDENSE GRANULE PROTEINS
650	1	4	_aGRA76
650	1	4	_aREPLICATION
650	1	4	_aTOXOPLASMA GONDII
650	1	4	_aVACCINE
650	1	4	_aVIRULENCE
700	1	2	_aZheng, X. N.
700	1	2	_aSun, L. X.
700	1	2	_aElsheikha, H. M.
700	1	2	_aLi, T. T.
700	1	2	_aGao, J.
700	1	2	_aWu, X. J.
700	1	2	_aWang, J. L.
856	4	0	_uhttps://drive.google.com/file/d/1Wmjju7roG8NGy0AgrIwHLcFZWGFzEB17/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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