establishment-and-culture-of-patient-derived-breast-organoids

Establishment and Culture of Patient-Derived Breast Organoids

Breast cancer is a complex disease that has been classified into several different histological and molecular subtypes. Patient-derived breast tumor ...

Cold Spring Harbor

Stony Brook University

The mammalian genome is pervasively transcribed and the functional significance of many long non-coding RNA (lncRNA) transcripts are gradually being elucidated.  () is one of the most well-studied lncRNAs.  is a highly conserved nuclear retained lncRNA that is abundantly expressed in cells and tissues and has been shown to play a role in regulating genes at both the transcriptional and post-transcriptional levels in a context-dependent manner. However,  has been shown to be dispensable for normal development and viability in mice. Interestingly, accumulating evidence suggests that  plays an important role in numerous diseases including cancer. Here, we discuss the current state-of-knowledge in regard to  with respect to its function, role in diseases, and the potential therapeutic opportunities for targeting  using antisense oligonucleotides and small molecules.

 Long Non-Coding RNA: Functional Implications.

The importance of studying the mechanistic aspects of long non-coding RNAs is being increasingly emphasized as more and more regulatory RNAs are being discovered. Non-coding RNA sequences directly associate with generic RNA-binding proteins as well as specific proteins, which cooperate in the downstream functions of the RNA and can also be dysregulated in various physiologic states and/or diseases. While current methods exist for identifying RNA-protein interactions, these methods require high quantities of input cells or use pooled capture reagents that may increase non-specific binding. We have developed a method to efficiently capture specific RNAs using less than one million input cells. One single oligonucleotide is used to pull down the target RNA of choice and oligonucleotide selection is driven by sequence accessibility. We perform thermal elution to specifically elute the target RNA and its associated proteins, which are identified by mass spectrometry. Ultimately, two target and control oligonucleotides are used to create an enrichment map of interacting proteins of interest.  eLife (2021), DOI: 10.7554/eLife.68263.

Disha Aggarwal

Cold Spring Harbor Laboratory

Cold Spring Harbor

Establishment and Culture of Patient-Derived Breast Organoids

Disha Aggarwal

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Cold Spring Harbor

Establishment and Culture of Patient-Derived Breast Organoids

Cold Spring Harbor Laboratory