Untitled Flashcards Set

High-throughput screens (HTS)

Subjecting many biological samples to simultaneous testing under given conditions, often using advanced machinery like microscopes and robotics.

How are we able to do HTS on mammalian cells?

cause of crispr. before we could only do it on yeast

RNA interference (RNAi)

A defense mechanism against viruses, mainly found in plants and some animals, that silences specific mRNA fragments in response to double-stranded RNA.

CRISPR

A genome-editing technology that uses the Cas9 protein to cut specific points in the DNA, allowing for targeted modifications at the genetic level.

dCas9 (dead Cas9)

An inactive form of Cas9 used in CRISPRi to block transcription without cutting DNA.

siRNA

Small interfering RNA, a form of RNA used in RNAi that silences genes by targeting mRNA.

shRNA

Short hairpin RNA that can be engineered into the genome to enable long-term gene silencing.

Genetic screening types

Includes silencing screens (like siRNA, shRNA, CRISPR-KO) and overexpression screens (like cDNAs, CRISPRa).

Arrayed screens

Type of screening where the contents of every plate are known, allowing immediate identification of the gene of interest.

Pooled screens

A screening method that examines a mixture of cells to identify genetic manipulations based on collective responses.

CRISPRa

A technique using CRISPR to enhance gene transcription by adding transcription factors while not blocking the promoter.

RISC complex

RNA-induced silencing complex that processes double-stranded RNA and degrades corresponding mRNA.

how do we use the RISC complex to silence genes?

The RISC complex utilizes small interfering RNA (siRNA) or microRNA (miRNA) to bind to target mRNA, leading to its degradation or translational repression, effectively silencing the gene.

Death associated proteins (DAP)

Proteins involved in programmed cell death that respond to signals like IFN-𝞬.

Microscopy in HTS

Utilized to analyze and measure cell division or other phenotypic changes during high-throughput screens. HTS is a method that allows for the rapid testing of thousands of compounds or conditions in biological assays.

cDNA libraries

Collections of complementary DNA used to study gene expression, typically requiring a wide range of gene sources.