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It has since been adopted for use as a tool in the genetic engineering of higher organisms. Designing an appropriate gRNA is an important element of genome editing with the CRISPR/Cas system. A gRNA can and at times does have unintended interactions ("off-targets") with other locations of the genome of interest.
CRISPR gene editing (CRISPR, pronounced / ˈ k r ɪ s p ə r / (crisper), refers to a clustered regularly interspaced short palindromic repeats") is a genetic engineering technique in molecular biology by which the genomes of living organisms may be modified.
A paper demonstrated that genome wide activation could be used to determine which proteins are involved in mediated resistance to a specific drug. [7] Another paper used genome wide activation of long, noncoding RNAs and observed that increasing the expression of certain long noncoding RNAs conferred resistance to the drug vemurafenib. [16]
When the modified organism mates and its DNA mixes with that of its mate, the CRISPR-Cas9 tool cuts the partner's DNA at the same spot where the gene drive is located in the first organism. The cell repairs the cut DNA by copying the gene drive from the first organism into the corresponding spot in the DNA of the offspring.
A transcriptional activator is a protein (transcription factor) that increases transcription of a gene or set of genes. [1] Activators are considered to have positive control over gene expression, as they function to promote gene transcription and, in some cases, are required for the transcription of genes to occur.
Targeted gene knockout using CRISPR/Cas9 requires the use of a delivery system to introduce the sgRNA and Cas9 into the cell. Although a number of different delivery systems are potentially available for CRISPR, [37] [38] genome-wide loss-of-function screens are predominantly carried out using third generation lentiviral vectors.
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Off-target genome editing refers to nonspecific and unintended genetic modifications that can arise through the use of engineered nuclease technologies such as: clustered, regularly interspaced, short palindromic repeats ()-Cas9, transcription activator-like effector nucleases (), meganucleases, and zinc finger nucleases (ZFN). [1]