基因表达差异显示实验

Electroporation of non-viral plasmid DNA is a valuable tool to alter gene expression in the adult central nervous system. It offers a number of advantages over viral gene delivery as non-viral plasmids can integrate larger inserts and reduce the risk of inducing unintended immunological r ...

Dendritic cells (DC) and macrophages (MΦ) play a pivotal role in antimicrobial defense, in the regulation of immune responses, and in maintaining tissue homeostasis. The analysis of DC and MΦ function relies on primary cells albeit these cells are known to be difficult to transfect. This makes the ...

Electroporation is a high-efficiency and low-toxicity physical gene transfer method. Traditional electroporation is limited to only low volume cell samples. Here we present a continuous cell electroporation method based on commonly used microfluidic chip fabrication techn ...

Due to their capacity for inducing strong and sequence specific gene silencing in cells, small interfering RNAs (siRNAs) are now recognized not only as powerful experimental tools for basic research in Molecular biology but with promising potentials in therapeutic development. Del ...

This chapter describes the in vivo delivery of conventional mRNA or alphaviral replicon RNA via intradermal electroporation. The use of RNA in clinical applications has several potential advantages compared to DNA. For instance, RNA cannot integrate into the host genome, and it does not co ...

Here, we outline a method for surface-mediated transfection of small interfering RNA (siRNA) using electroporation. Cells are cultured on an electrode carrying siRNA. After the cells are adhered on the electrode surface, an electric pulse is applied to release siRNA from the surface and tra ...

Electroporation-mediated gene transfer (electro-transfection) is a powerful tool to introduce nucleic acid compounds such as plasmid DNAs, antisense oligonucleotides, and short interfering RNAs (siRNAs) into the cells. Electro-transfection is a physical gene transfer me ...

Short interfering RNAs (siRNAs) represent new potential therapeutic tools owing to their capacity to induce strong, sequence-specific, gene silencing in cells. Electropulsation is one of the physical methods successfully used to transfer siRNA into living cells in vitro and in vivo. A ...

Electroporation serves as an attractive nonviral gene delivery approach for its effectiveness, operational simplicity, and no restrictions of probe or cell type. The commercial electroporation systems have been widely adopted in research and clinics with protocols usually co ...

Electroporation is an effective physical delivery method. A variety of factors have been shown to affect the electroporation-mediated gene delivery efficiency. Here we report the usefulness of noncoding short-fragment DNA (sf-DNA) for facilitating electroporation-media ...

Cell transfection efficiency often determines the success of cell-based gene therapy. Cell transfection via Nucleofector technology yields high transfection efficiency and low cytotoxicity. However, owing to trade secrecy, the components in each buffer are unknown, which not o ...

Electrical pulses directly and effectively boost both in vitro and in vivo gene transfer, but this process is greatly affected by non-electrical factors that exist during electroporation. These factors include, but are not limited to, the types of cells or tissues used, property of DNA, DNA for ...

Membrane electropermeabilization is the observation that the permeability of a cell membrane can be transiently increased when a micro-millisecond external electric field pulse is applied on a cell suspension or on a tissue. Applicative aspects for the transfer of foreign molecules ...

Homing endonucleases (HEs) are natural enzymes that cleave long DNA target with a high specificity and trigger homologous recombination at the exact site of the break. Such mechanisms can thus be used for all the applications covered today by the generic name of “genome engineering”: targeted ...

Homing endonucleases and other site-specific endonucleases have potential applications in genome editing, yet efficient targeting requires a thorough understanding of DNA-sequence specificity. Here, we describe a modified two-plasmid genetic selection in Escherichia ...

Homing endonucleases recognize long DNA sequences and generate site-specific DNA double-stranded breaks. They can serve as a powerful genomic modification tool in various industrial and biomedical applications. Here, we describe a two-plasmid bacterial selection system for c ...

Homing endonuclease I-CreII has been used to study the consequences and repair of a double-strand break (DSB) in the chloroplast genome of Chlamydomonas and Arabidopsis. Since I-CreII is from a mobile psbA intron of Chlamydomonas, it cleaves the psbA gene of an intronless-psbA strain of Chlam ...

5′RLM-RACE is a PCR-based technique used to map the 5′ termini of transcripts in both eukaryotic and prokaryotic organisms. Free-standing homing endonuclease promoters often lack recognizable promoters making predicting the transcriptional start site challenging. Further ...

Mapping the precise position of endonucleolytic cleavage sites is a fundamental experimental technique used to describe the function of a homing endonuclease. However, these proteins are often recalcitrant to cloning and over-expression in biological systems because of toxic ...

Fungal mitochondrial genomes act as “reservoirs” for homing endonucleases. These enzymes with their DNA site-specific cleavage activities are attractive tools for genome editing and gene therapy applications. Bioprospecting and characterization of naturally occurring ...