Mouse In Vivo Placental Targeted CRISPR Manipulation

Annemarie J Carver; Robert J Taylor; Hanna E Stevens

doi:10.3791/64760

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Mouse In Vivo Placental Targeted CRISPR Manipulation

Journal article

Open access

Peer reviewed

Mouse In Vivo Placental Targeted CRISPR Manipulation

Annemarie J Carver, Robert J Taylor and Hanna E Stevens

Journal of visualized experiments, Vol.194, e64760

04/14/2023

DOI: 10.3791/64760

PMCID: PMC10664715

PMID: 37125793

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https://pmc.ncbi.nlm.nih.gov/articles/PMC10664715/pdf/nihms-1943174.pdfView

Open Access

Abstract

The placenta is an essential organ that regulates and maintains mammalian development in utero. The placenta is responsible for the transfer of nutrients and waste between the mother and fetus and the production and delivery of growth factors and hormones. Placental genetic manipulations in mice are critical for understanding the placenta's specific role in prenatal development. Placental-specific Cre-expressing transgenic mice have varying effectiveness, and other methods for placental gene manipulation can be useful alternatives. This paper describes a technique to directly alter placental gene expression using CRISPR gene manipulation, which can be used to modify the expression of targeted genes. Using a relatively advanced surgical approach, pregnant dams undergo a laparotomy on embryonic day 12.5 (E12.5), and a CRISPR plasmid is delivered by a glass micropipette into the individual placentas. The plasmid is immediately electroporated after each injection. After dam recovery, the placentas and embryos can continue development until assessment at a later time point. The evaluation of the placenta and offspring after the use of this technique can determine the role of time-specific placental function in development. This type of manipulation will allow for a better understanding of how placental genetics and function impact fetal growth and development in multiple disease contexts.

Fetus

Pregnancy

Animals

Clustered Regularly Interspaced Short Palindromic Repeats

Female

Fetal Development

Mammals

Mice

Placenta - metabolism

Details

Title: Subtitle: Mouse In Vivo Placental Targeted CRISPR Manipulation
Creators: Annemarie J Carver
Robert J Taylor
Hanna E Stevens
Resource Type: Journal article
Publication Details: Journal of visualized experiments, Vol.194, e64760
DOI: 10.3791/64760
PMID: 37125793
PMCID: PMC10664715
NLM abbreviation: J Vis Exp
eISSN: 1940-087X
Language: English
Date published: 04/14/2023
Academic Unit: Psychiatry; Iowa Neuroscience Institute
Record Identifier: 9984400640302771

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