An investigator designed a crRNA to target mouse myostatin gene with sequence TGACGATTATCACGCTACCA. Please very how many places in the genome does this sequence target if mismatch number of 2 or less is allowed (assuming DNA bulge and RNA Bulge size is 0)? 3 2 4 1
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- If all members of a transposable element family can besilenced by dsRNA synthesized from a single familymember, how is it possible for one element family (likeTc1) to have 32 copies in the C. elegans genome whileanother family (Tc2) has fewer than 5 copies?One problem that researchers sometimes encounterwhen editing genomes with CRISPR/Cas9 is that oneor more loci other than the intended target can berecognized by Cas9/sgRNA and cleaved. Part of thereason is that single base pair mismatches betweenthe target site and the sgRNA in the 5′-most half of the 20 bp DNA/RNA hybrid do not prevent Cas9cleavage of the target site. How could scientists usebioinformatics to avoid such off-target effects?One complication of making a transgenic animal is that thetransgene might integrate at random into the coding region, orthe regulatory region, of an endogenous gene. What might be theconsequences of such random integrations?
- Bacteriophage lambda is as one of the routinely used molecular cloning vectors, which alsoserved as a model system for the study of bacteriophage morphogenesis, DNA replication, andgene regulation.a) With the aid of a diagram, generally narrate how a foreign gene can be inserted into alambda insertion vector and subsequently infect an Escherichia coli cell.b) You are cloning a 7.5 kb gene into a lambda gt10 vector, utilizing a restriction site whichspecifically present in the vector. State the restriction site that you can use for this purposeand suggest a screening procedure to indicate successful integration of the gene into thehost genome.For the sake of simplicity, Fig. 10.4 omitted one stepof cDNA library construction. The figure impliedthat the last step of the process is the ligation ofblunt-ended cDNAs into plasmid cloning vectors.Although such ligation reactions can occur, in realitythey are highly inefficient. Instead, scientists convertblunt-ended cDNA molecules into sticky-ended molecules using adapters, and then they ligate thecDNAs into vectors with compatible sticky ends.Adapters are short, partly double-stranded DNAmolecules made by hybridization of two singlestranded oligonucleotides made in a DNA synthesizer.Suppose that the following two oligonucleotides weresynthesized and then mixed together at high concentration and at a temperature that promotes hybridization of complementary DNA sequences:5′ CCCCCG 3′5′ AATTCGGGGG 3′a. Draw the hybridized DNA molecules. These arethe adapters.b. Suppose you added the adapters and ligase enzymeto blunt-ended cDNAs at a very high molar ratio ofadapters to cDNAs, so…siRNA and CRISPR/Cas9 each interfere with gene expression by a sequence-directed mechanism. A. Identify the target macromolecule (DNA, RNA or Protein) of a specific siRNA/RISC andCRISPR/Cas9 complexsiRNA:CRISPR/Cas9: B. If a scientist would like to permanently turn off expression of a particular human gene inthe liver for the lifetime of an individual, which of these techniques would bepreferable? Briefly explain why
- Locate a gene expression data set on Gene Expression Omnibus using "expression by hybridization" involving Rabies virus or related virus Use GEO2R to perform pairwise group analysis Explore and identify the most regulated genes: gene/protein functions of Rabies virusand Define the significance of these genes/proteins in the disease process.A certain cDNA of size 2 kb hybridized to eight genomicfragments of total size 30 kb and contained two shortESTs. The ESTs were also found in two of the genomicfragments each of size 2 kb. Sketch a possible explanation for these resultsThe ability to selectively modify the genome in the mouse has revolutionized mouse genetics. Outline the procedure for generating a knockout mouse at a specific genetic locus. How can the loxP-Cre system be used to conditionally knock out a gene? What is an important medical application of knockout mice?
- 70Pre-initiation begins with either TFIIF or TATA binding protein binding to the core promoter. Yesorno 71Exons tend to be non-conserved during evolution, whereas introns usually are conserved. Yesorno 72Transfer RNA molecules fold into L-shaped three-dimensional structures. YesornoBefore the technology existed to synthesize RNAmolecules of defined sequence like those in Fig. 8.5,similar experiments were performed with syntheticmRNAs of undefined sequence. For example, RNAsconsisting only of Us and Gs could be synthesized invitro, but they would have random sequences. Supposea pool of random-sequence RNAs was synthesized ina reaction mixture containing three times as much UTPas GTP, and that the resulting RNAs were translatedin vitro.a. How many different codons exist in the RNAs?b. How many different amino acids would you find inthe polypeptides synthesized?c. Why are your answers to (a) and (b) not the same?d. How often would you expect to find each of thecodons in (a)?e. In what proportions would you expect to find eachof the amino acids in the polypeptides?f. If you did this experiment—that is, synthesizedrandom-sequence RNAs containing a 3:1 ratio ofU:G, and quantified the amount of each amino acidin the polypeptides produced—prior to knowledgeof the genetic…Refer to the DNA sequence provided: 3’ -TACTGAAGCGGCAGCCCCGCATGAGTAGACCTTACT-5’ a. What is the mRNA transcript of the anticoding strand of the DNA model? b. What is the amino acid sequence of the polypeptide chain that will be translated from the mRNA in (a)?