BAd-CRISPR: Inducible gene knockout in interscapular brown adipose tissue of adult mice

Romanelli, Steven M.; Lewis, Kenneth T.; Nishii, Akira; Rupp, Alan C.; Li, Ziru; Mori, Hiroyuki; Schill, Rebecca L.; Learman, Brian S.; Rhodes, Christopher J.; MacDougald, Ormond A.

doi:10.1016/j.jbc.2021.101402

Cited by 9 publications

(2 citation statements)

References 85 publications

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“…CRISPR Cas9 has assisted the expansion of knowledge on the cellular and molecular biology of different cell types. Romanelli et al, [54] developed a transgenic mouse line producing the Cas9 proteín within brown adipocytes. The authors implemented gene editing in brown mouse adipocytes to analyze the function of various active genes in such cells.…”

Section: Crispr Cas9 Based Technology For Cell Biologymentioning

confidence: 99%

Genetic Editing with CRISPR Cas9: recent Biomedical and Biotechnological Applications

Garzón Posse,

Pinilla Peña,

Rivas Velásquez

et al. 2024

Universitas Scientiarum

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The use of a novel and powerful technology that allows for the precise editing of the genetic material of various organisms is becoming widespread. This technology derives from bacterial and archaeal defense machinery and is called CRISPR Cas9. Unlike other gene editing tools that exclusively rely on proteins, CRISPR Cas9 utilizes interactions between the target DNA and an RNA sequence that guides the Cas9 enzyme to alter the structure of a target gene. Various genome locations can be edited thanks to the ease of programming different guide RNA sequences, facilitating its use and implementation. Furthermore, the non-active version of the Cas9 protein, guided by its corresponding RNA, can be utilized for visualization processes of genetic material or, more recently, for the regulation of the transcription process. Considering the recent advances and possibilities in biomedical and biotechnological research, we must understand that the exploration of this technology is just beginning, and its eventual applications will influence the world around us on multiple levels. In this review, we describe the biological foundations of the functioning of the Cas9 nuclease, together with selected applications of its use in editing and regulating specific sections of the genetic material of various organisms. We also discuss some bioethical issues surrounding this subject.

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Section: Crispr Cas9 Based Technology For Cell Biologymentioning

confidence: 99%

Genetic Editing with CRISPR Cas9: recent Biomedical and Biotechnological Applications

Garzón Posse,

Pinilla Peña,

Rivas Velásquez

et al. 2024

Universitas Scientiarum

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“…This protocol leverages key technique advances towards the uses of AAVs to target adipose tissues and the methods to inject interscapular brown adipose tissue (iBAT). 2 , 3 , 4 , 5 , 6 , 7 , 8 …”

Section: Before You Beginmentioning

confidence: 99%

Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo

Xue¹,

Wu²,

Qiu³

2022

STAR Protocols

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N^G,N^G‐Dimethylarginine Dimethylaminohydrolase 1 Expression Is Dispensable for Cold‐ or Diet‐Induced Thermogenesis

Park,

Lee,

Bagon

et al. 2024

Advanced Biology

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The strategy to activate thermogenic adipocytes has therapeutic potential to overcome obesity as they dissipate surplus energy as heat through various mechanisms. NG,NG‐dimethylarginine dimethylaminohydrolases (DDAHs) are enzymes involved in the nitric oxide‐protein kinase G signaling axis which increases thermogenic gene expression. However, the role of DDAHs in thermogenic adipocytes has not been elucidated. The adipocyte‐specific Ddah1 knockout mice are generated by crossing Ddah1fl/fl mice with adiponectin Cre recombinase mice. Adipocyte‐specific DDAH1 overexpressing mice are generated using adeno‐associated virus‐double‐floxed inverse open reading frame (AAV‐DIO) system. These mice are analyzed under basal, cold exposure, or high‐fat diet (HFD) conditions. Primary inguinal white adipose tissue cells from adipocyte‐specific Ddah1 knockout mice expressed comparable amounts of Ucp1 mRNA. Adipocyte‐specific DDAH1 overexpressing mice do not exhibit enhanced activation of thermogenic adipocytes. In addition, when these mice are exposed to cold environment or fed an HFD, their body temperature/weight and thermogenesis‐related gene and protein expressions are unchanged. These findings indicate that DDAH1 does not play a role in either cold‐ or diet‐induced thermogenesis. Therefore, adipocyte targeting DDAH1 gene therapy for the treatment of obesity is unlikely to be effective.

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BAd-CRISPR: Inducible gene knockout in interscapular brown adipose tissue of adult mice

Cited by 9 publications

References 85 publications

Genetic Editing with CRISPR Cas9: recent Biomedical and Biotechnological Applications

Genetic Editing with CRISPR Cas9: recent Biomedical and Biotechnological Applications

Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo

N^G,N^G‐Dimethylarginine Dimethylaminohydrolase 1 Expression Is Dispensable for Cold‐ or Diet‐Induced Thermogenesis

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BAd-CRISPR: Inducible gene knockout in interscapular brown adipose tissue of adult mice

Cited by 9 publications

References 85 publications

Genetic Editing with CRISPR Cas9: recent Biomedical and Biotechnological Applications

Genetic Editing with CRISPR Cas9: recent Biomedical and Biotechnological Applications

Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo

NG,NG‐Dimethylarginine Dimethylaminohydrolase 1 Expression Is Dispensable for Cold‐ or Diet‐Induced Thermogenesis

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N^G,N^G‐Dimethylarginine Dimethylaminohydrolase 1 Expression Is Dispensable for Cold‐ or Diet‐Induced Thermogenesis