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Cleavage and Blastulation

Cleavage and Blastulation

After a large-single-celled zygote is produced via fertilization, the process of cleavage occurs while zygotes travel through the uterine tube. Cleavage is a mitotic cell division that does not result in growth. With each round of successive cell division, daughter cells get increasingly smaller.

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Related Experiment Video

Updated: Jul 11, 2025

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Published on: January 7, 2020

Pri-miRNA cleavage assays for the Microprocessor complex.

Thi Nhu-Y Le¹, Cong Truc Le¹, Tuan Anh Nguyen¹

¹Division of Life Science, The Hong Kong University of Science & Technology, Hong Kong, P.R. China.

Methods in Enzymology

|November 4, 2023

Summary

This summary is machine-generated.

The Microprocessor complex (MP) is essential for microRNA (miRNA) production in mammals. This study details an in vitro assay to analyze MP

Keywords:

DGCR8 DROSHA Microprocessor Pri-miRNA in vitro pri-miRNA cleavage miRNA

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Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

The Microprocessor complex (MP) is critical for microRNA (miRNA) biogenesis in animals, initiating miRNA production by cleaving primary miRNAs (pri-miRNAs).
Accurate MP enzymatic activity is essential for correct miRNA sequencing, expression, and cellular function.
Various RNA elements and cofactors within pri-miRNAs can modulate MP cleavage activity, influencing miRNA output.

Purpose of the Study:

To establish and describe an in vitro pri-miRNA cleavage assay for evaluating Microprocessor complex (MP) activity.
To provide a method for assessing how different RNA substrates and conditions affect MP cleavage efficiency.
To enable the analysis of MP enzymatic activity, adaptable for high-throughput screening.

Main Methods:

Purification of the human Microprocessor complex (MP) from HEK293E cells.
In vitro transcription to synthesize various pri-miRNA substrates.
Conducting in vitro pri-miRNA cleavage assays using purified MP and synthesized substrates.

Main Results:

The study successfully established a reproducible in vitro assay for assessing MP cleavage activity.
The described methodology allows for the evaluation of MP function with diverse RNA substrates.
The assay utilizes standard laboratory equipment and reagents, making it accessible for various research settings.

Conclusions:

The developed in vitro pri-miRNA cleavage assay is a valuable tool for studying Microprocessor complex (MP) function in miRNA biogenesis.
This assay can be optimized for high-throughput analysis, facilitating the screening of numerous RNA substrates and conditions.
Understanding MP activity is crucial for deciphering miRNA regulation and its impact on cellular processes.