RNA Gene Silencing

RNA Gene Silencing

With over 20 years experience in custom synthesis for the biomedical research communities, BSI has developed the expertise to deliver custom synthesized RNA with quality that meets all your RNAi, siRNA, shRNA and other RNA projects.

RNA Extraction Kits

These kits are optimized for use with 10,000-2,000,000 cells as starting material, providing 2-25 µg of total RNA for microarray hybridizations and other life sciences applications. Use these kits in conjunction with ArrayIt MiniAmp mRNA Amplification Kit and Indirect Amino Allyl Labeling Products.

Common RNA

Before the nucleotides are linked together, they exist separately as ribonucleoside triphosphates (NTPs). As shown below, the NTPs contain one of the four common RNA bases, A, C, G, and U, linked to a five-carbon ribose sugar, linked, in turn, to a chain of three phosphate groups. During RNA synthesis, a covalent, "phosphodiester" bond is formed between one of the three phosphate groups on one NTP and a hydroxyl group on another. The two other phosphate groups that were part of the original NTP are released.


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Protein Biosynthesis

Protein Biosynthesis (synthesis) is the process in which cells build proteins. The term is sometimes used to refer only to protein translation but more often it refers to a multi-step process, beginning with amino acid synthesis and transcription which are then used for translation. Protein biosynthesis, although very similar, differs between prokaryotes and eukaryotes.

Amino acid synthesis
Amino acids are the monomers which are polymerized to produce proteins. Amino acid synthesis is the set of biochemical processes (metabolic pathways) which build the amino acids from carbon sources like glucose. Not all amino acids may be synthesised by every organism, for example adult humans have to obtain 8 of the 20 amino acids from their diet.

Transcription
Transcription is the process by which an mRNA template, encoding the sequence of the protein in the form of a trinucleotide code, is transcribed from the genome to provide a template for translation. Transcription copies the template from one strand of the DNA double helix, called the template strand.Transcription can be divided into 3 stages: Initiation, Elongation and Termination, each regulated by a large number of proteins such as transcription factors and coactivators that ensure the correct gene is transcribed in response to appropriate signals.The DNA strand is read in the 3' to 5' direction and the mRNA is transcribed in the 5' to 3' direction by the RNA polymerase.

Translation
The synthesis of proteins is known as translation. Translation occurs in the cytoplasm where the ribosomes are located. Ribosomes are made of a small and large subunit which surrounds the mRNA. In translation, messenger RNA (mRNA) is decoded to produce a specific polypeptide according to the rules specified by the genetic code. This uses an mRNA sequence as a template to guide the synthesis of a chain of amino acids that form a protein. Translation is necessarily preceded by transcription. Translation proceeds in four phases: activation, initiation, elongation and termination (all describing the growth of the amino acid chain, or polypeptide that is the product of translation).In activation, the correct amino acid (AA) is joined to the correct transfer RNA (tRNA). While this is not technically a step in translation, it is required for translation to proceed. The AA is joined by its carboxyl group to the 3' OH of the tRNA by an ester bond.

Antisense Technology

At Bio-Synthesis, we offer discovery scale synthesis to mid-scale multi gram quantities for clinical diagnostic applications, in addition to the synthesis of these modified oligos, we routinely assist customers in the design of the oligos that are particularly suited to their applications.

Antisense therapy
Antisense therapy is a form of treatment for genetic disorders or infections. When the genetic sequence of a particular gene is known to be causative of a particular disease, it is possible to synthesize a strand of nucleic acid (DNA, RNA or a chemical analogue) that will bind to the messenger RNA (mRNA) produced by that gene and inactivate it, effectively turning that gene "off". This is because mRNA has to be single stranded for it to be translated. The goal of antisense applications is to shut down activity of a defined gene by blocking transcription or translation within cells. Oligonucleotides for antisense assays must be nuclease resistant against cellular nucleases, must be able to cross cellular membranes and must inherit both high binding affinity and specificity for the target sequence. In many cases, they also must have the ability to induce RNase H cleavage.

Antisense DNA
Antisense molecules interact with complementary strands of nucleic acids, modifying expression of genes. Some regions within a double strand of DNA code for genes, which are usually instructions specifying the order of amino acids in a protein along with regulatory sequences, splicing sites, non coding introns and other complicating details. For a cell to use this information, one strand of the DNA serves as a template for the synthesis of a complementary strand of RNA. The stability of the RNA-DNA duplex in term of hybrization and half-life is crucial to successful gene inhibition. Vigorous research activity in the area of nucleic acid chemistry has been devoted in developing novel base analgos that are resistant to degradation and that possess strong hybridization properties. This includes the classical phosphorothioate linkages, propyne analogs and the latest lock nucleic acid (LNA) base analogs, and peptide nucleic acid (PNA).

Antisense mRNA
Antisense mRNA is an mRNA transcript that is complementary to endogenous mRNA. In other words, it is a non-coding strand complementary to the coding sequence of mRNA; this is similar to negative-sense viral RNA. Introducing a transgenic coding for antisense mRNA is a technique used to block expression of a gene of interest. Radioactively-labeled antisense mRNA can be used to show the level of transcription of genes in various cell types. Some alternative antisense structural types are being experimentally applied as antisense therapy, with at least one antisense therapy approved for use in humans.

RNA Synthesis

With over 20 years experience in custom synthesis for the biomedical research communities, Biosyn has developed the expertise to deliver custom synthesized RNA with quality that meets all your RNAi, siRNA, sh RNA and other RNA projects.

RNA Synthesis
RNA synthesis is said to proceed in the 5′ to 3′ direction, reflecting the fact that the attachment of new nucleotides always occurs at the 3′ hydroxyl group of the growing RNA chain. RNA synthesis goes through phases that are typical of polymerization processes: initiation, elongation, and termination, yielding an RNA product of defined size and sequence.

Common RNA
Before the nucleotides are linked together, they exist separately as ribonucleoside tripolyphosphate (NTPs). As shown below, the NTP's contain one of the four common RNA bases, A, C, G, and U, linked to a five-carbon ribosome sugar, linked, in turn, to a chain of three phosphate groups. During RNA synthesis, a covalent, "phosphorescent" bond is formed between one of the three phosphate groups on one NTP and a hydroxyl group on another. The two other phosphate groups that were part of the original NTP are released.

Nucleotides
RNA, like DNA, is a polymer of nucleotides. Each nucleotide consists of a sugar that is attached to a phosphate group and any one of four bases. The RNA polymerase, as it builds the chain of nucleotides, processes only one of the two complementary strands of DNA. This DNA strand is referred to as the template strand. The least confusing name for the other DNA strand is "the non-template strand.