The RNAi Consortium

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The RNAi Consortium, or TRC, is a public-private partnership whose mission is to create libraries of small hairpin RNAs (shRNAs) for 15'000 human and 15'000 mouse genes. These libraries should help the scientific community to analyse gene function by RNAi. The consortium is based at the Broad Institute of the MIT and Harvard University, and includes 6 MIT- and Harvard-associated institutions and 5 international life sciences organizations. Verified RNAi clones and entire libraries are made available both by Sigma-Aldrich and Open Biosystems.

Gene Basic physical and functional unit of heredity

In biology, a gene is a sequence of nucleotides in DNA or RNA that codes for a molecule that has a function. During gene expression, the DNA is first copied into RNA. The RNA can be directly functional or be the intermediate template for a protein that performs a function. The transmission of genes to an organism's offspring is the basis of the inheritance of phenotypic trait. These genes make up different DNA sequences called genotypes. Genotypes along with environmental and developmental factors determine what the phenotypes will be. Most biological traits are under the influence of polygenes as well as gene–environment interactions. Some genetic traits are instantly visible, such as eye color or number of limbs, and some are not, such as blood type, risk for specific diseases, or the thousands of basic biochemical processes that constitute life.

Broad Institute biomedical and genomic research center

The Eli and Edythe L. Broad Institute of MIT and Harvard, often referred to as the Broad Institute, is a biomedical and genomic research center located in Cambridge, Massachusetts, United States. The institute is independently governed and supported as a 501(c)(3) nonprofit research organization under the name Broad Institute Inc., and is partners with Massachusetts Institute of Technology, Harvard University, and the five Harvard teaching hospitals.

Harvard University Private research university in Cambridge, Massachusetts, United States

Harvard University is a private Ivy League research university in Cambridge, Massachusetts, with about 6,700 undergraduate students and about 15,250 postgraduate students. Established in 1636 and named for its first benefactor, clergyman John Harvard, Harvard is the United States' oldest institution of higher learning. Its history, influence, wealth, and academic reputation have made it one of the most prestigious universities in the world.

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Hairpin selection

A set of candidate hairpins are selected based on the 1st Refseq transcript from each NCBI gene. They should be 21mers, be at least 25bp from start of the coding sequence and no closer than 150bp from its end. Candidates are scored based on various empirical rules (see the Broad Institute's web site for a complete list ) and then BLASTed against 2 transcriptome sets. Hairpins that are unique for a Unigene cluster and a RefSeq NM identifier are preferred. Lastly, the candidates are spaced to have 1 hairpin in the 3' untranslated region and 4 in the coding sequence.

National Center for Biotechnology Information database arm of the US National Library of Medicine

The National Center for Biotechnology Information (NCBI) is part of the United States National Library of Medicine (NLM), a branch of the National Institutes of Health (NIH). The NCBI is located in Bethesda, Maryland and was founded in 1988 through legislation sponsored by Senator Claude Pepper.

Transcriptome set of all RNA molecules in one cell or a population of cells

The transcriptome is the set of all RNA molecules in one cell or a population of cells. It is sometimes used to refer to all RNAs, or just mRNA, depending on the particular experiment. It differs from the exome in that it includes only those RNA molecules found in a specified cell population, and usually includes the amount or concentration of each RNA molecule in addition to the molecular identities.

Hairpin vector

Selected hairpins are cloned into the vector pLKO1, which is a multipurpose plasmid that can be propagated in bacteria, transfected into mammalian cell lines or used for generation of lentiviruses. It contains resistance genes against ampicillin and puromycin.

Viral vectors are tools commonly used by molecular biologists to deliver genetic material into cells. This process can be performed inside a living organism or in cell culture. Viruses have evolved specialized molecular mechanisms to efficiently transport their genomes inside the cells they infect. Delivery of genes, or other genetic material, by a vector is termed transduction and the infected cells are described as transduced. Molecular biologists first harnessed this machinery in the 1970s. Paul Berg used a modified SV40 virus containing DNA from the bacteriophage λ to infect monkey kidney cells maintained in culture.

Plasmid small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently

A plasmid is a small DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria; however, plasmids are sometimes present in archaea and eukaryotic organisms. In nature, plasmids often carry genes that benefit the survival of the organism, such as by providing antibiotic resistance. While the chromosomes are big and contain all the essential genetic information for living under normal conditions, plasmids usually are very small and contain only additional genes that may be useful in certain situations or conditions. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. In the laboratory, plasmids may be introduced into a cell via transformation.

Bacteria A domain of prokaryotes – single celled organisms without a nucleus

Bacteria are a type of biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, bacteria have a number of shapes, ranging from spheres to rods and spirals. Bacteria were among the first life forms to appear on Earth, and are present in most of its habitats. Bacteria inhabit soil, water, acidic hot springs, radioactive waste, and the deep portions of Earth's crust. Bacteria also live in symbiotic and parasitic relationships with plants and animals. Most bacteria have not been characterised, and only about 27 percent of the bacterial phyla have species that can be grown in the laboratory . The study of bacteria is known as bacteriology, a branch of microbiology.

Release

Release 1 of the TRC lentiviral shRNA libraries consist of about 35'000 shRNA constructs against 5300 human (25'000 clones) and 2200 mouse genes (10'000 clones). Release 2 of the human shRNA library contained an additional 9'500 clones. Releases occur roughly every quarter. The completing of both mouse and human libraries is envisioned for the end of 2006 or beginning of 2007.

Related Research Articles

Complementary DNA Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary dna, not circular DNA, not c-DNA) is used in a variety of molecular cloning experiments as well as serving as

In genetics, complementary DNA (cDNA) is DNA synthesized from a single-stranded RNA template in a reaction catalyzed by the enzyme reverse transcriptase. cDNA is often used to clone eukaryotic genes in prokaryotes. When scientists want to express a specific protein in a cell that does not normally express that protein, they will transfer the cDNA that codes for the protein to the recipient cell. cDNA is also produced naturally by retroviruses and then integrated into the host's genome, where it creates a provirus.

In genetics, an expressed sequence tag (EST) is a short sub-sequence of a cDNA sequence. ESTs may be used to identify gene transcripts, and are instrumental in gene discovery and in gene-sequence determination. The identification of ESTs has proceeded rapidly, with approximately 74.2 million ESTs now available in public databases.

This is a list of topics in molecular biology. See also index of biochemistry articles.

Short hairpin RNA

A short hairpin RNA or small hairpin RNA is an artificial RNA molecule with a tight hairpin turn that can be used to silence target gene expression via RNA interference (RNAi). Expression of shRNA in cells is typically accomplished by delivery of plasmids or through viral or bacterial vectors. shRNA is an advantageous mediator of RNAi in that it has a relatively low rate of degradation and turnover. However, it requires use of an expression vector, which has the potential to cause side effects in medicinal applications.

Kv1.1 protein-coding gene in the species Homo sapiens

Potassium voltage-gated channel subfamily A member 1 also known as Kv1.1 is a shaker related voltage-gated potassium channel that in humans is encoded by the KCNA1 gene. The Isaacs syndrome is a result of an autoimmune reaction against the Kv1.1 ion channel.

RNA silencing or RNA interference refers to a family of gene silencing effects by which gene expression is negatively regulated by non-coding RNAs such as microRNAs. RNA silencing may also be defined as sequence-specific regulation of gene expression triggered by double-stranded RNA (dsRNA). RNA silencing mechanisms are highly conserved in most eukaryotes. The most common and well-studied example is RNA interference (RNAi), in which endogenously expressed microRNA (miRNA) or exogenously derived small interfering RNA (siRNA) induces the degradation of complementary messenger RNA. Other classes of small RNA have been identified, including piwi-interacting RNA (piRNA) and its subspecies repeat associated small interfering RNA (rasiRNA).

mir-148/mir-152 microRNA precursor family

In molecular biology, miR-148 is a microRNA whose expression has been demonstrated in human, mouse, rat and zebrafish. miR-148 has also been predicted in chicken.

DNA polymerase beta protein-coding gene in the species Homo sapiens

DNA polymerase, beta, also known as POLB, is an enzyme present in eukaryotes. In humans, it is encoded by the POLB gene.

HIC2 protein-coding gene in the species Homo sapiens

Hypermethylated in cancer 2 protein is a protein that in humans is encoded by the HIC2 gene.

ZNF365 protein-coding gene in the species Homo sapiens

Protein ZNF365 is a protein that in humans is encoded by the ZNF365 gene.

Dharmacon Inc., now known as Dharmacon, a Horizon Discovery Group company, was founded in 1995 by Stephen Scaringe as Dharmacon Research to develop and commercialize a new technology for RNA oligonucleotide synthesis. The original Dharmacon focus and vision was to develop 2'-ACE RNA technology as the standard for RNA synthesis and to advance RNA oligo-dependent applications and technologies. When RNA interference (RNAi) emerged in the late 1990s, Dharmacon was poised to provide RNAi-related products to the multitude of academic and industry researchers. Dharmacon has become an important resource for those investigating the mechanisms of siRNA -induced gene knockdown and applying the specificity and potency of RNAi to human biotherapeutics. Dharmacon's expertise in bioinformatics, RNA biology, and synthesis chemistry has allowed it to develop a complete line of products for the RNAi researcher.

IMAGE cDNA clones are a collection of DNA vectors containing cDNAs from various organisms including human, mouse, rat, non-human primates, zebrafish, pufferfish, Xenopus (frogs), and cow. Together they represent a more or less complete set of expressed genes from these organisms. IMAGE stands for integrated molecular analysis of genomes and their expression.

Chloride channel accessory 3, also known as CLCA3, is a protein which in humans is encoded by the CLCA3P pseudogene. The protein encoded by this gene is a chloride channel. According to the HGNC, this protein is not expressed in humans but is in certain other species such as mouse. However, some conflicting reports state that human cells produce and glycosylate this protein.

Knockout rat

A knockout rat is a genetically engineered rat with a single gene turned off through a targeted mutation used for academic and pharmaceutical research. Knockout rats can mimic human diseases and are important tools for studying gene function and for drug discovery and development. The production of knockout rats was not economically or technically feasible until 2008.

HIKESHI protein-coding gene in the species Homo sapiens

HIKESHI is a protein important in lung and multicellular organismal development that, in humans, is encoded by the HIKESHI gene. HIKESHI is found on chromosome 11 in humans and chromosome 7 in mice. Similar sequences (orthologs) are found in most animal and fungal species. The mouse homolog, lethal gene on chromosome 7 Rinchik 6 protein is encoded by the l7Rn6 gene. When the l7Rn6 protein is disrupted in mice, the mice display severe emphysema at birth as a result of disorganization of the Golgi apparatus and formation of aberrant vesicular structures within club cells.

Complementarity (molecular biology) describes a relationship between two structures each following the lock-and-key principle

In molecular biology, complementarity describes a relationship between two structures each following the lock-and-key principle. In nature complementarity is the base principle of DNA replication and transcription as it is a property shared between two DNA or RNA sequences, such that when they are aligned antiparallel to each other, the nucleotide bases at each position in the sequences will be complementary, much like looking in the mirror and seeing the reverse of things. This complementary base pairing allows cells to copy information from one generation to another and even find and repair damage to the information stored in the sequences.

CAPRIN2 protein-coding gene in the species Homo sapiens

caprin family member 2, also known as CAPRIN2, is a human gene.

Benitec Biopharma Ltd ASX: BLT is an Australian biotechnology company founded in 1997. It is engaged in the development of gene-silencing therapies for the treatment of chronic and life-threatening diseases using DNA-directed RNA interference (ddRNAi) technology.

DNA-directed RNA interference

DNA-directed RNA interference (ddRNAi) is a gene-silencing technique that utilizes DNA constructs to activate an animal cell’s endogenous RNA interference (RNAi) pathways. DNA constructs are designed to express self-complementary double-stranded RNAs, typically short-hairpin RNAs (shRNA), that once processed bring about silencing of a target gene or genes. Any RNA, including endogenous mRNAs or viral RNAs, can be silenced by designing constructs to express double-stranded RNA complementary to the desired mRNA target.

Gregory James Hannon is a professor of molecular cancer biology and director of the Cancer Research UK Cambridge Institute at the University of Cambridge. He is a Fellow of Trinity College, Cambridge while also serving as a director of cancer genomics at the New York Genome Center and an adjunct professor at Cold Spring Harbor Laboratory.