CRISPR/Cas9-based gene editing in genetic studies have the following attractive feature(s) in defining functions of genes related to human health? OA Multiplex CRISPR can be designed to simultaneously knock out two or several functionally redundant genes. B. Designing a guide RNA based on the DNA sequence in the 3'-end of a protein-coding gene is more efficient than all other regions. Oc Cas9 causes a double-stranded DNA break immediately before the NGG PAM site. O D.CRISPR/Cas9 knockout has no off-target concern.
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- What advantages do cDNA libraries provide over genomic DNA libraries? Describe cloning applications where the use of a genomic library is necessary to provide information that a cDNA library cannot.The following illustrates a jagged double-strand DNA break resulting from Cas9 cleavage that occurred in the first step of genome editing using CRISPR-Cas9 technology: 5'-GCGCCGTCC 3'-CGCGGC CTGTCAGGCGACACT-3' AGGGACAGTCCGCTGTGA-5' Which of the double-stranded DNA sequences listed below (A-D) is expected to result from repair of the break above via non-homologous end joining? Note: this question is not asking what kinds of mutations result from NHEJ repair of Cas9 cleavage in general, but specifically what is expected to result from repair of the jagged cut illustrated above? In the answer choices below, sequences that are the same in all four options are shown in bold to help you spot the differences. A. 5'-GCGCCGCTGTCAGGCGACACT-3' 3'-CGCGGCGACAGTCCGCTGTGA-5' B. 5'-GCGCCGTCTGTCAGGCGACACT-3 3'-CGCGGCAGACAGTCCGCTGTGA-5' C. 5'-GCGCCGTCCCTGTCAGGCGACACT-3' 3'-CGCGGCAGGGACAGTCCGCTGTGA-5' D. 5'-GCGCCGAGACTGTCAGGCGACACT-3' 3'-CGCGGCTCTGACAGTCCGCTGTGA-5'1. You are investigating a protein that has the amino acid sequence N ... Ala – Thr - Asn – Trp – Lys - Arg - Gly – Phe – Thr ... C within its primary structure. You found that several of the mutations affecting this protein produced shortened protein molecules that terminated within this region. In one of the mutants, the Asn became the terminal (last) amino acid. (a) What DNA single-base changes(s) would cause the protein to terminate at the Asn residue? (b) What other potential sites do you see in the DNA sequence encoding this protein where mutation of a single base pair would cause premature termination of translation? >
- Which of the following statements is not true about the guide RNA (see this interactive demonstration for help with this question: https://www.biointeractive.org/classroom-resources/crispr-cas9-mechanism-applications) Group of answer choices it contains a sequence of 20 nucleotides that matches a specific sequence in a cell’s DNA When the guide RNA is combined with Cas9, it will guide Cas9 to the target sequence its target sequence can be almost any sequence as long as it occurs near a PAM motif it is a nuclease, a type of enzyme that cleaves DNAWhich of the following is a correct statement about CRISPR-Cas-9 gene editing? Group of answer choices A single guide RNA (sgRNA) recognizes a genomic region followed by 5'-NGG-3' PAM sequence A single guide RNA (sgRNA) recognizes a genomic region followed by a long sequence palindrome repeat A single guide DNA (sgDNA) recognizes a genomic region followed by 5'-NGG-3' PAM sequence PAM sequences induce single stranded breaks that are then repaired by the CAS-9 enzymeWhy is genome editing by CRISPR-Cas advantageous over traditionalmethods for creating knockout or transgenic animals?Explain your answers.
- CRISPR/Cas9 mediated genome editing has provided an unparalleled means of manipulating the genome. Describe in 2 sentences/lines maximum what three requirements must be met in order to target a specific dsDNA break at a given site in genome of the target organism using this system?a gm animal that may be approved for human consumption by the time this book is published is a super muscly pig made by inactivation of the myostatin gene. During normal development, myostatin protein prevents the overgrowth of muscles. (Given that effective deletions can be made in the genome by using CRISPR/Cas9. It targets deletions of varied length. By this technique, the transcriptional status of the target gene is not affected. The gene deletions produced by CRISPR/Cas9 lead to the production of the correct type of junctions at very high frequencies), how could the super muscly pig have been generated? Answer the questions in the photo as well. ( i included my answer for 22a in the question above, if you believe you have a better answer by all means include that as well)both noncoding and coding). a. Which types of SNPs affect protein production or function for the gene of interest? b. Which types of SNPs might be identified in a GWAS? Biolnteractive.org (including Updated November 20% Page 1 of
- You are working for a pharmaceutical company are are tasked with creating E. Coli that express an influenza antigen (a protein that is recognized by our immune system) fortreatment of the influenza virus. The cDNA of the antigen is available in a carrier vector(pCARRY) that contains a kanamycin resistance gene as shown on the left below. Inorder to get E. Coli to express the antigen, you need to clone the gene for the influenzaantigen into an expression vector that contains an E. Coli-specific promoter. You obtain such an expression vector, which contains an ampicillin resistance gene as shown on the right below (pBACTERIA) a. An end generated by digestion with BamH1 can be ligated to an end generated by digestion with BlgII. Why is this possible? b. To clone the gene for the influenza antigen from pCARRY into pBACTERIA plasmid:i. What restriction enzyme should you use to digest pCARRY? ii. What restriction enzyme should you use to digest pBACTERIA? c. After you ligate the products of…. Early gene-cloning experiments involved insertion at one restriction site in the vector; for example, the insert would have an EcoRI site at each end, and the vector would be opened at an RI site prior to ligation. Under what circumstances would asymmetric cloning be desirable, with the inset having a different restriction site at each endCRISPR-Cas9 provides an unprecedented means of modifying the genome of almost any organism. Which of the following component(s) of this system contribute to the specificity of the endonucleolytic cleavage Options: the sequence of the sgRNA the PAM sequence Cas9 a and b a, b and c