A nonviral method for gene transfer into mammalian cells has been developed using physical force which accelerates plasmid DNA-coated gold particles to high -speed and penetrate the mammalian cells. This technology of gene transfer via a biolistic transfection method has been shown to have multiple applications to mammalian gene transfer systems. This method has also been adapted for delivery of other macromolecules like RNA, microRNA, and proteins. A broad range of somatic cell types, including primary cell cultures and established cell lines, have been successfully transfected ex vivo or in vitro by using the gene gun technology, either as suspension or adherent cells in cultures. This chapter describes the general procedures for in vitro DNA transfection by particle-mediated delivery to nonadherent and adherent cells. These procedures can be readily employed by using the Helios gene gun system (Bio-Rad, Hercules, CA) based on the Accell design.
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http://dx.doi.org/10.1007/978-1-62703-110-3_11 | DOI Listing |
Genome Biol Evol
January 2025
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
The human malaria parasite Plasmodium falciparum evolved from a parasite that infects gorillas, termed Plasmodium praefalciparum. The sialic acids on glycans on the surface of erythrocytes differ between humans and other apes. It has recently been shown that the P.
View Article and Find Full Text PDFSci Rep
January 2025
Department of Gynecology and Obstetrics, Yijishan Hospital of Wannan Medical College, Wuhu, Anhui, People's Republic of China.
The ABCB4 gene encodes multidrug resistance protein 3(MDR3), which is a phosphatidylcholine(PC) transfer enzyme that transfers lecithin from the inner part of the phospholipid bilayer to the extracellular bile. The occurrence of intrahepatic cholestasis of pregnancy(ICP) is closely related to ABCB4 variants, but there is limited research on this topic in southern Anhui, China. We sequenced ABCB4 in pregnant women with ICP and healthy pregnant women to explore the relationship.
View Article and Find Full Text PDFNat Commun
January 2025
Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA, USA.
While all native tRNAs undergo extensive post-transcriptional modifications as a mechanism to regulate gene expression, mapping these modifications remains challenging. The critical barrier is the difficulty of readthrough of modifications by reverse transcriptases (RTs). Here we use Induro-a new group-II intron-encoded RT-to map and quantify genome-wide tRNA modifications in Induro-tRNAseq.
View Article and Find Full Text PDFArch Biochem Biophys
January 2025
Department of Nuclear Medicine, Hefei BOE Hospital, Hefei City, Anhui Province, 241000, China. Electronic address:
Background: Diffuse large B-cell lymphoma (DLBCL) is a prevalent and aggressive form of non-Hodgkin's lymphoma with a complex etiology. NOP2/Sun domain 2 (NSUN2) is an RNA methyltransferase that has been linked to the regulation of gene expression in various cancers. However, the function of NSUN2 in DLBCL, specifically its contribution to exosome-driven tumor progression, remains to be thoroughly elucidated.
View Article and Find Full Text PDFFree Radic Biol Med
January 2025
Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide-CSIC-JA, Sevilla, Spain; CIBERER, U729, Instituto de Salud Carlos III, Madrid, Spain. Electronic address:
The interference of the expression of each of the genes involved in the synthesis of coenzyme Q (CoQ) in Drosophila melanogaster can help to understand the pathophysiology of CoQ-dependent mitochondrial diseases in humans. We have knocked-down all genes involved in the CoQ biosynthesis pathway at different temperatures to induce depletion of CoQ at different levels throughout the body and in a tissue-specific manner. The efficiency of the knockdowns was quantified by Q-RTPCR and determination of CoQ levels by HPLC-UV+ECD.
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