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- The amount of DNA inintronic regionof genes can be grwater than the amount of DNA in exons. trueWhat is the role of the following molecules in the CRISPR mechanism? * Sequence of 3 nucleotide Not a RNA significant sequence Endonuclease motif that is Host genetic synthesized role in information to match a a binding CRISPR target site mechanism sequence Cas9 guide RNA DNA polymerase PAM Cellular DNA TRNA O O 口 ロ 口 ロロGene targeting and genome editing are both techniques for removingor modifying a particular gene, each of which can produce thesame ultimate goal. Describe some of the differences between theexperimental methods used for these two techniques.
- Nonhomologous end-joining (NHEJ) of a doublestrand break almost always results in perfect resealingof the DNA lesion, without the loss or gain of nucleotide pairs. Yet CRISPR/Cas9, which produces doublestrand breaks, is a highly efficient method of makingsmall deletions or insertions at the targeted site. Howcan you resolve this apparent contradiction?In a typical microbiology laboratory, reasons for no bands from a gel of a polymerase chain reaction may bedue to errors relating to omission of ingredients in the reaction mix and absence of the target sequence inthe template DNA. Based on (i) primer problem and (ii) purity/potential contamination of the target sequence, explain the reason for non-appereance on bands.Ehat primer sets could be amplify the following DNA sequence? AATACGTCGCATGGggatccttttttatgcatg
- R plasmid R determinant CRISPR tracrRNA sgRNA M III III III A CRISPR nuclease that can cut foreign DNA and destroy it. Small circle of extragenomic DNA that contains antibiotic resistant genes. Move a new gene into a genome. Antibiotic resistant genes. Trans-acting ('tracer") CRISPR RNA segment as a stem-loop structure that binds to Cas and gRNA. Interfere with mRNA of a gene so that it cannot be expressed. Remove a gene from a genome.Transcribe and translate the following DNA sequence (nontemplate strand): 5’-ATGGCCGGTTATTAAGCA-3’Label the following parts of the CRISPR-Cas 9 system. C A B C D Target DNA strand PAM sequence Cas9 protein Single-guide RNA
- The recognition sequence for the restriction enzyme Sau3AIis GATC; in the recognition sequence forthe enzyme BamHI—GGATCC—the four internal bases areidentical to the Sau3AI sequence. The single-stranded endsproduced by the two enzymes are identical. Suppose youhave a cloning vector that contains a BamHI recognitionsequence and you also have foreign DNA that was cut withSau3AI. Can this DNA be ligated into the BamHI site of the vector,and if so, why?You are cloning the genome of a new DNA virus into pUC18.You plate out your transformants on ampicillin plates containing X-gal and pick one blue colony and one white colony.When you check the size of the inserts in each plasmid (blueand white), you are surprised to fi nd that the plasmid fromthe blue colony contains a very small insert of approximately60 bp, while the plasmid from the white colony does notappear to contain any insert at all. Explain these results.The schematic dlagram below shows the functional organization of transcribing RNA polymerase. Match parts of the diagram (labeled A-G) with the corresponding term from the answer list (design ated 1-12). Polymerase movement ANA POLYMERASE A Rowinding of DNA Unwinding of DNA E NTPs Note that some of the items from the answer list should NOT be used. Labels E and F point to the specific ends of DNA/RNA molecules. 1. Template strand 2. RNA transcript 3. Replication origin 4 Non-Template strand 5. Leading strand 6. 3 end 7. Primer 8. Replication fork 9. Transcription bubble 10. 5' end 11. DNA RNA nybrid 12. Lagging strand