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The new antiretroviral therapy for human immunodeficiency virus can slow viral replication but is unable to eliminate HIV infection because proviral DNA embedded in the host genome remains genetically silent in reservoir cells and is replication-competent on interruption or cessation of ART. We developed a novel HIV gene therapy using synthetic guide RNA/Cas9-ribonucleoprotein as a non-viral formulation in this research. In lymphoid and monocytic latent HIV cell lines, we developed a line of gRNAs targeting various HIV genes critical for HIV replication and testing their antiviral stability and cellular cytotoxicity. HIV-gRNA/Cas9 RNP-treated cells showed effective viral suppression with no apparent cytotoxicity, as shown by the significant reduction of latent HIV DNA reactivation and RNA replication in comparison to scramble gRNA control. Moreover, HIV-gRNA/Cas9 RNP instutive gen expression, reduced infectious viral particle production, and produced specific DNA cleavages in the targeted HIV genes that can be determined by DNA sequencing.
Source link: https://doi.org/10.3390/v14091902
Abstract CRISPR/Cas9 has dramatically changed how we do genetic analysis by giving us a tool for precise sequence editing. However, new CRISPR/Cas9 applications have appeared that do not require nuclease use. Kuhl et al. , a leading role for Ctf19 in kinetochore-derived meiotic recombination suppression in a key role for Ctf19 in kinetochore-mediated recombination suppression, according to the accompanying paper. Kuhl et al. explains that Kuhl et al. Using the power of CRISPR/Cas9 to bind specific genomic sequences, Kuhl et al. explains. CRISPR/Cas9 is a graduate student who receives undergraduate students with background on chromosomes, meiosis, recombination, and CRISPR/Cas9 to enhance their understanding of Kuhl et al.
Source link: https://doi.org/10.1093/genetics/iyac109
Vitelline membrane proteins are the key proteins that make up insect eggs's inner shell and are a vital component of egg formation and embryogenesis. PxVMP26 was specifically expressed in female adults and was highly expressed in the ovary, according to both qPCR and western blot tests. The expression level in the ovarian tube with an incomplete yolk was significantly higher than that in the ovarian tube with a complete yolk, according to a new anatomical report. There was no significant difference between the number of eggs laid within three days between wild and mutant individuals, but a lower egg hatchability was found. Using CRISPR/Cas9 technologies, this first review of the role of the VMP gene in P. xylostella's oocyte differentiation and embryonic development provides a basis for testing new genetic control targets of P. xylostella.
Source link: https://doi.org/10.3390/ijms23179538
We investigated for the presence of pre-existing adaptive immune responses to their respective Cas9 homologs, SaCas9 and SpCas9, based on the fact that these two bacterial species cause infections in the human race at high rates. We tested for the presence of anti-Cas9 antibodies using human serum and were able to find antibodies against both, with 79% of donors staining against SaCas9 and 69% of donors staining against SpCas9. In 46 percent of donors, we discovered anti-SaCas9 T-cells against the two homologs in human peripheral blood. The CRISPR-Cas9 system in human trials shows that there are pre-existing humoral and cell-mediated adaptive immune responses to Cas9 in humans, a factor that must be considered as the CRISPR-Cas9 system enters clinical trials.
Source link: https://doi.org/10.1101/243345
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