1. karaj islamic azad university 2. karaj azad university
Abstract
Introduction
Molecular genetic studies in cell and animal models have evolved our knowledge of discovering causal mechanisms relating molecular events to phenotypes. recently, the bacterial system, crispr (clustered regularly-interspaced palindromic repeats) harnessed to facilitate genetic manipulations. it is challenging to elucidate the roles of these complex combinations in genetic alterations on tumor evolution largely due to the difficulty of creating transgenic animals with multiple perturbed alleles. thus, we aim to review the instruction process of multi-gene deficient in mice models having lung cancer
Methods
kras oncogene and the tumor suppressor genes p53 and lkb1 (stk11), are the top of three most frequently mutated genes in lung adenocarcinoma in human. to modeling of the dynamics of mutations, a single vector made, that’s capable of generating krasg12d mutations while simultaneously knocking out p53 and lkb1 genes. then adeno-associated vector (aav) encompass related sgrnas was transected into a cre-dependent cas9 mice. to modeling of a missense gain-of-function kras mutation, an hdr donor template designed
Results
Four weeks later, lungs were harvested from mice and indels of p53 and lkb1 were identified at the predicted cutting site, with 0.1% p53 indels and 0.4% lkb1 indels in the whole lung. the frequency of indels in p53 and lkb1 and targeted krasg12d mutations was increased over time. this was potentially due to the selective growth advantage of mutant cells
Conclusion
Collectively, these data suggest that delivery of vector into the lungs of cre-dependent cas9 mice generates the krasg12d mutation and putative loss-of-function mutations in p53 and lkb1 genes and also can have the potential of crispr tool in precise modeling of variety of biological process and disease
