What is transient transfection and the steps

Posted by beauty33 on August 27th, 2021

Transient transfection is one of the ways to introduce DNA into eukaryotic cells. In transient transfection, recombinant DNA is introduced into a highly infectious cell line to obtain temporary but high-level expression of the target gene. The transfected DNA does not need to be integrated into the host chromosome, the transfected cells can be harvested in a shorter time than stable transfection, and the expression of the target gene in the lysate can be detected? 

Features

The transient expression method has many advantages:

Transient transfection is one of the ways to introduce DNA into eukaryotic cells. In transient transfection, recombinant DNA is introduced into a highly infectious cell line to obtain temporary but high-level expression of the target gene. The transfected DNA does not need to be integrated into the host chromosome, the transfected cells can be harvested in a shorter time than stable transfection, and the expression of the target gene in the lysate can be detected.

First, the operation is simple and the cycle is short. The traditional stable transformation system requires at least several months or even longer time from transformation to obtaining transgenic plants, and the transformation efficiency is relatively low. The transient expression system only needs 2 to 4 weeks, avoiding the tedious process of plant tissue culture (Kapila et al. al., 1997);

Second, the expression efficiency is high. When the single-stranded T-DNA enters the plant cell, not only the genes transformed into the plant genome can be expressed, but many free foreign genes that are not integrated into the plant genome can also be expressed with a higher copy number. Its expression is not affected by gene location and gene silencing, so the expression efficiency is higher, and it can reach several times the expression level of stable transformation (Janssenand Gardner, 1990; Kapila et al., 1997; Yang et al., 2000; Wroblewski et al ., 2005);

Third, it is safe and efficient. Stably expressed transgenic plants have food chain pollution, gene drift caused by pollen transmission and other transgenic safety issues (Twyan et al., 2003). Gene transfer and its expression in transient expression are an independent process and do not produce heritable offspring. Its biological safety is much higher, and the test results are more intuitive and reliable;

Fourth, there is no need to screen genes. Stable transgenic transformation usually requires preliminary screening of antibiotics or other screening genes. The transgenic plants also have relative transgenic safety issues. Transient expression does not require screening of genes, which reduces the unsafe factors caused by antibiotics in transgenes, and the vector can be smaller. , It is easier to transform larger target fragments and multiple genes;

Fifth, it can be performed on whole plants. When analyzing the interaction of exogenous gene expression and biotic or abiotic stress, it can more truly reflect the gene expression pattern in plants.