CRISPR-cas9 is a powerful technique for gene editing. Please place the stages of a CRISPR-cas9 gene editing workflow in the correct order from stages 1-9. Note, there is one incorrect, redundant answer. - Add green fluorescent protein gene sequence into plasmid to aid selection of correctly transfected cells.- Clonal characterisation (analysis of phenotype).- Synthesize DNA insert oligonucleotides.- Select for correctly gene edited cells (using antibiotic resistance and/or color production for example).- Transfect cells- Purify plasmids- Clone into CRISPR-cas9 expression vector.- Design a set of targeting sequences using algorithms.- Clonal isolation.- Sequence.

CRISPR-cas9 is a powerful technique for gene editing. Please place the stages of a CRISPR-cas9 gene editing workflow in the correct order from stages 1-9. Note, there is one incorrect, redundant answer. - Add green fluorescent protein gene sequence into plasmid to aid selection of correctly transfected cells.- Clonal characterisation (analysis of phenotype).- Synthesize DNA insert oligonucleotides.- Select for correctly gene edited cells (using antibiotic resistance and/or color production for example).- Transfect cells- Purify plasmids- Clone into CRISPR-cas9 expression vector.- Design a set of targeting sequences using algorithms.- Clonal isolation.- Sequence.

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 8QP

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Please place the stages of a CRISPR-cas9 gene editing workflow in the correct order below. Note, there is one incorrect and state why? Sequence Select for correctly gene edited cells (using antibiotic resistance and/or colour production for example) Clonal isolation Synthesise DNA insert oligonucleotides Clonal characterisation (analysis of phenotype) Add green fluorescent protein gene sequence into plasmid to aid selection of correctly transfected cells. Transfect cells Clone into CRISPR-cas9 expression vector Purify plasmids Design a set of targeting sequences using algorithms.
Place the stages of a CRISPR-cas9 gene editing workflow in the correct order (there is one incorrect stage) Design a set of targeting sequences using algorithms. Clonal isolation Transfect cells Synthesise DNA insert oligonucleotides Purify plasmids Sequence Clonal characterisation (analysis of phenotype) Select for correctly gene edited cells (using antibiotic resistance and/or colour production for example) Add green fluorescent protein gene sequence into plasmid to aid selection of correctly transfected cells. Clone into CRISPR-cas9 expression vector
What is the function of gRNA in CRISPR-Cas technologies?It cuts DNA at a targeted sequence and inserts a new sequence -.It cuts protein at a targeted sequence and inserts a new sequence.It ensures the Cas9 enzyme cuts at the correct place in the DNA.It ensures the Cas9 enzyme cuts at the correct place in the RNA.It replaces uracil nucleotides with thymine nucleotides.
You are performing an experiment using CRISPR-cas9 to genetically modify the LacZ gene of a culture of E. coli. After you run the experiment, you decide to use gel electrophoresis to genotype the different bacterial cultures to determine if the gene editing was successful. How could your electrophoresis results confirm that the PCR was successful? And how could your electrophoresis results confirm that you successfully extracted genomic DNA from your bacterial samples?
There may be more than one correct answer, select all that apply. To clone genes, a plasmid vector should contain a(n) a.) telomeres b.) orgin of replication c.) antibiotic resistance gene d.) restriction endonuclease cut site The CRISPR/Cas system has revolutionized the efficiency at which scientists can experimentally induce a mutation in a gene to investigate the functional role of the protein it encodes. Which of the following are required in the CRISPR/Cas system? Cas9 nuclease Bacterial genome Access to target gene Guide RNA complementary to gene of interest
With reference to the image below, discuss the process and principle involved for screening/selection of hosts (last stage of cloning) containing the intended recombinant plasmid. LacZ' = Gene for alpha-peptide of β-galactosidase.
In order to construct a genome library, which statement most accurately describes the correct order of steps in the process? Fragment chromosomal DNA, digest plasmid vector, ligate genomic DNA and digested plasmid vector, transform ligation into bacteria, select for plasmid marker Fragment chromosomal DNA and plasmid vector, ligate genomic DNA and plasmid vector, transform ligation into bacteria, select for gene marker Fragment chromosomal DNA and plasmid vector, ligate genomic DNA and plasmid vector, transform ligation into bacteria, select for plasmid marker Fragment chromosomal DNA, ligate genomic DNA and plasmid vector, transform ligation into bacteria, select for plasmid marker
What are the major components of the CRISPR-Cas9 system? What mechanism does it employ to combine DNA? Explain the process of how the CRISP-Cas9 system is able to create recombinant DNA. Relate the idea of gene modification to the fields of vaccines and applied microbiology as well.
Consider the following human genetic diseases: hemophilia, Down syndrome, cystic fibrosis, and brain cancer. Which are the best candidates for treatment with CRISPR-Cas genome editing, and which have the largest hurdles to overcome? Why?
When using a conventional plasmid cloning vector containing a b-galactosidase gene, it is possible to perform a "blue-white screen" to determine which bacteria have taken up a plasmid into which a DNA fragment as been inserted, as opposed to those that have taken up just reclosed plasmid vector, by growing the transformed cells on nutrient agar plates containing the artificial b-gal substrate X-gal. Will bacteria that have taken up a plasmid into which a DNA fragment has been inserted form a blue colony or a white colony when grown on this medium? Briefly explain why these bacteria would form a colony of the color you chose.
Why is genome editing by CRISPR-Cas advantageous over traditionalmethods for creating knockout or transgenic animals?Explain your answers.
CRISPR/Cas9 can be used in genome editing. Among the following statements, which one is correct? The CRISPR and the Cas9 parts both recognise the DNA target sequence and then recruit an endonuclease for cutting it The Cas9 part recognises the DNA target sequence and the CRISPR part cuts it The CRISPR and the Cas9 parts both recognise and cut the DNA target sequence. The CRISPR part recognises the DNA target sequence and Cas9 cuts it.