yruvate dehydrogenase kinase-4 (PDK-4) is a widely expressed isoform of PDK and is responsible for inhibiting pyruvate ehydrogenase (PDH) complex activity via phosphorylation. When PDK-4 is active, the PDH complex is in its phosphorylated ate and pyruvate cannot be converted into acetyl-CoA for entry into the citric acid cycle. Instead, in gluconeogenic tissues like ver and kidney, pyruvate is channeled to support gluconeogenesis. Conversely, when PDK-4 is inactive, the PDH complex eadily converts pyruvate into acetyl-CoA. researcher wants to determine the role of PDK-4 on blood glucose homeostasis. To accomplish this, she proposes to generate PDK-4 knockout mouse using CRISPR technology. To create the knockout mouse, she must first generate PDK-4 knockout mbryos to implant into pseudopregnant female mice. rrange the steps to outline the process by which she will generate the PDK-4 knockout embryos. First step Last step Answer Bank Generate Cas9 mRNA and sgRNA specific to the target sequence using in vitro transcription. Isolate single cell mouse embryos and inject them with Cas9 mRNA and sgRNA. Identify a protospacer-adjacent motif (PAM) adjacent to PDK-4 in the genome. Determine the DNA sequence of the region of the genome containing PDK-4.
yruvate dehydrogenase kinase-4 (PDK-4) is a widely expressed isoform of PDK and is responsible for inhibiting pyruvate ehydrogenase (PDH) complex activity via phosphorylation. When PDK-4 is active, the PDH complex is in its phosphorylated ate and pyruvate cannot be converted into acetyl-CoA for entry into the citric acid cycle. Instead, in gluconeogenic tissues like ver and kidney, pyruvate is channeled to support gluconeogenesis. Conversely, when PDK-4 is inactive, the PDH complex eadily converts pyruvate into acetyl-CoA. researcher wants to determine the role of PDK-4 on blood glucose homeostasis. To accomplish this, she proposes to generate PDK-4 knockout mouse using CRISPR technology. To create the knockout mouse, she must first generate PDK-4 knockout mbryos to implant into pseudopregnant female mice. rrange the steps to outline the process by which she will generate the PDK-4 knockout embryos. First step Last step Answer Bank Generate Cas9 mRNA and sgRNA specific to the target sequence using in vitro transcription. Isolate single cell mouse embryos and inject them with Cas9 mRNA and sgRNA. Identify a protospacer-adjacent motif (PAM) adjacent to PDK-4 in the genome. Determine the DNA sequence of the region of the genome containing PDK-4.
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Transcribed Image Text:Pyruvate dehydrogenase kinase-4 (PDK-4) is a widely expressed isoform of PDK and is responsible for inhibiting pyruvate
dehydrogenase (PDH) complex activity via phosphorylation. When PDK-4 is active, the PDH complex is in its phosphorylated
state and pyruvate cannot be converted into acetyl-CoA for entry into the citric acid cycle. Instead, in gluconeogenic tissues like
liver and kidney, pyruvate is channeled to support gluconeogenesis. Conversely, when PDK-4 is inactive, the PDH complex
readily converts pyruvate into acetyl-CoA.
A researcher wants to determine the role of PDK-4 on blood glucose homeostasis. To accomplish this, she proposes to generate
a PDK-4 knockout mouse using CRISPR technology. To create the knockout mouse, she must first generate PDK-4 knockout
embryos to implant into pseudopregnant female mice.
Arrange the steps to outline the process by which she will generate the PDK-4 knockout embryos.
First step
Last step
Answer Bank
Generate Cas9 mRNA and sgRNA specific to the target sequence using in vitro transcription.
Isolate single cell mouse embryos and inject them with Cas9 mRNA and sgRNA.
Identify a protospacer-adjacent motif (PAM) adjacent to PDK-4 in the genome.
Determine the DNA sequence of the region of the genome containing PDK-4.
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