UC Riverside

Since the discovery of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats), it has allowed for genome editing to be utilized in a way never seen before. Although it can create knockout and point mutations at ease, it can still be difficult to create knockin mutations. In order to combat this issue, GENEWRITE was created utilizing the reverse transcription function of the LINE-1 (long interspersed element-1) and combining it with the commonly used Cas (CRISPR associated proteins). This also worked around the need for CRISPR to silence NHEJ as it can be utilized in the protocol as well. This, however, made it difficult for GENEWRITE to function in organisms such as prokaryotes and archaea which did not already carry a NHEJ pathway. To identify prokaryotes and archaea the GENEWRITE protocol can function in, a bioinformatics analysis was done to identify the known forms of NHEJ in prokaryotes and archaea. Additionally, it was recently found that TnpB, a transposon associated gene from the IS200/IS605 family found in bacteria, is a RNA-guided DNA endonuclease that could be programmed similar to Cas proteins. To verify its potential function within the transposon, a quantitative microscopy study was carried out to observe how TnpB affected transposition in real tim