Explain why each of the criteria listed below are important in selection of a guide RNA (hint: look under Instructions on CHOPCHOP). Under what conditions do these criteria decrease the efficiency of binding of guide RNA to its target sequence (e.g. does more, or less, self-complementarity decrease efficiency)? a) % GC (= # of G and C’s in the guide RNA sequence divided by 20 – the length of the guide RNA; this may also be called GC content) b) Self-complementarity: c) Off-Targets (0, 1, 2, 3). In your answer, explain what the numbers 0, 1, 2, 3 indicate. For example - Is it better to have off-target sites or no off-target sites? Are off-targets in the 1 column more or less likely to decrease efficiency than off-targets in the 3 column?
DNA and RNA
Deoxyribonucleic acid (DNA) is usually called the blueprint of life. Deoxyribose is a monosaccharide that has a key function in the synthesis of deoxyribonucleic acid. One less oxygen-containing hydroxyl group occurs in deoxyribose sugar. Nucleic acid, deoxyribonucleic acid, is one of the natural components. Deoxyribonucleic acid is a double-stranded molecule. Watson and Crick postulated the double-stranded model of the helix. A deoxyribonucleic acid is a molecular group that carries and transmits genetic information from parents to offspring. All eukaryotic and prokaryotic cells are involved.
DNA as the Genetic Material
DNA, or deoxyribonucleic acid, is a long polymeric nucleic acid molecule discovered in the late 1930s. It is a polymer; a long chain-like molecule made up of several monomers connected in a sequence. It possesses certain characteristics that qualify it as a genetic component. Certain organisms have different types of nucleic acids as their genetic material - DNA or RNA.
Genetics
The significant branch in science which involves the study of genes, gene variations, and the organism's heredity is known as genetics. It is also used to study the involvement of a gene or set of genes in the health of an individual and how it prevents several diseases in a human being. Thus, genetics also creates an understanding of various medical conditions.
DNA Replication
The mechanism by which deoxyribonucleic acid (DNA) is capable of producing an exact copy of its own is defined as DNA replication. The DNA molecules utilize a semiconservative method for replication.
- Explain why each of the criteria listed below are important in selection of a guide RNA (hint: look under Instructions on CHOPCHOP). Under what conditions do these criteria decrease the efficiency of binding of guide RNA to its target sequence (e.g. does more, or less, self-complementarity decrease efficiency)?
a) % GC (= # of G and C’s in the guide RNA sequence divided by 20 – the length of the guide RNA; this may also be called GC content)
b) Self-complementarity:
c)
Off-Targets (0, 1, 2, 3). In your answer, explain what the numbers 0, 1, 2, 3 indicate. For example - Is it better to have off-target sites or no off-target sites? Are off-targets in the 1 column more or less likely to decrease efficiency than off-targets in the 3 column?
CHOPCHOP is a program designed for CRISPR design. The software allows users to input sequences, select desired specificity or sgRNA length, and optimize for low GC content.
Step by step
Solved in 2 steps
