遗传学实验技术

Ex vivo activated B cells are an alternative source of antigen presenting cells (APC). However, the ability of ex vivo activated B cells to function as potent APCs has been a concern especially when compared to dendritic cells (DC). Herein, we introduce a strategy to modulate antigen presentation a ...

mRNA-based transfection is an attractive strategy for manipulation of gene expression for gain-of-function studies and therapeutic applications. As a potential therapeutic regulator, mRNA transfection has mainly been hampered by poor delivery strategies, combined with la ...

Appropriate gene delivery systems are essential for successful gene therapy in clinical medicine. Cationic lipid-mediated delivery is an alternative to viral vector-mediated gene delivery where transient gene expression is desirable. However, cationic lipid-mediated de ...

The translational efficiency and stability of synthetic mRNA in both cultured cells and whole animals can be improved by incorporation of modified cap structures at the 5′-end. mRNAs are synthesized in vitro by a phage RNA polymerase transcribing a plasmid containing the mRNA sequence in the ...

In vitro transcription of DNA with phage RNA polymerases is currently the most efficient method to produce long sequence-specific RNA. While the reaction can yield large quantities of RNA, it contains impurities due to various unwanted activities of the polymerases. Here, we described an e ...

The in vitro synthesis of long RNA can be accomplished using phage RNA polymerase and template DNA. However, the in vitro synthesized RNA, unlike those transcribed in vivo in cells, lacks nucleoside modifications. Introducing modified nucleosides into in vitro transcripts is important ...

mRNA has become an important alternative to DNA as a tool for cell reprogramming. To be expressed, exogenous DNA must be transmitted through the cell cytoplasm and placed into the nucleus. In contrast, exogenous mRNA simply has to be delivered into the cytoplasm. This can result in a highly uniform tr ...

The import of functional nucleic acids like messenger RNA into mammalian cells has proven to be a powerful tool in cell biology and several delivery systems have been described. However, as targeting of particular cell types is a major challenge and RNA vaccination represents a promising means ...

Retrovirus-mediated mRNA transfer (RMT) combines the advantageous features of retrovirus-mediated cell targeting and entry with the controlled transfer of mRNAs. We have recently exploited this strategy for the dose-controlled transfer of recombinases and DNA transposas ...

Nuclear transplantation has been used to study genomic imprinting. Available nuclear transfer methods include pronuclear transfer (PNT), intracytoplasmic sperm injection, and round spermatid injection. By generating uniparental embryos that have exclusively patern ...

Pluripotent stem cells can provide us with an enormous cell source for in vitro model systems for development. In 2006, new methodology was designed to generate pluripotent stem cells directly from somatic cells, and these cells were named induced pluripotent stem cells (iPSCs). This method ...

Chromosomes acquire different epigenetic marks during oogenesis and spermatogenesis. After fertilization, if retained and selected, these differences may result in imprinting effects. Rather than being an oddity, imprinting effects have been found in many sexually reprodu ...

Genomic imprinting is an epigenetic phenomenon that results in the silencing of alleles, dependent on their parent of origin. Within vertebrates, this phenomenon is restricted only to the mammals and has been identified in eutherians and marsupials but not in the egg-laying monotremes. M ...

Computational studies on imprinted genes can have very different purposes: one major aim of these studies is the identification of DNA elements that distinguish imprinted genes from biallelically expressed genes. Comparative studies may help to identify imprinting regulatory e ...

Plants are excellent systems for discovering and studying epigenetic phenomena, such as transposon silencing, RNAi, imprinting, and DNA methylation. Imprinting, referring to preferential expression of maternal or paternal alleles, plays an important role in reproduction de ...

It is now estimated that the human genome encodes thousands of long noncoding (lnc)RNAs. These novel molecules are causing a paradigm shift in the field of molecular biology as a number of lncRNAs have been shown to be involved in a wide range of biological functions including regulation of gene expr ...

Next-generation sequencing of noncoding RNA (ncRNA) libraries has become an essential tool for the profiling of ncRNAs and the identification of novel ncRNA species. Here, we describe the generation of a ncRNA-derived complementary DNA (cDNA) library by 3′-tailing of ncRNAs by CTP and poly ...

Researchers whose experimental models are mammalian oocytes and preimplantation embryos are often limited by the yield of nucleic acids that can be isolated from such a small sample size. In addition, the limited number of cells from these types of samples makes the simultaneous recovery of R ...

Methylated DNA immunoprecipitation (MeDIP) is an immunocapturing approach for unbiased enrichment of DNA that is methylated on cytosines. The principle is that genomic DNA is randomly sheared by sonication and immunoprecipitated with an antibody that specifically recognizes ...

Development of high-throughput sequencing technologies now enables genome-wide analysis of DNA methylation of mammalian cells and tissues. Here, we present a protocol for Reduced Representation Bisulfite Sequencing (RRBS) applicable to low amounts of starting material (fr ...