Presenter
Henry Purnell
Document Type
Poster
Publication Date
2024
Abstract
Due to its small genome and dominant haploid life stage, Physcomitrium patens (formerly Physcomitrella patens) has become a model organism for studying plant genetics. The focus of this study was the Cellulose Synthase gene superfamily of P.patens, specifically the Cellulose Synthase-Like Ds (CSLD) family of genes. Previous studies into CSLD genes were preformed using RNA interference (RNAi) to preform a loss of function analysis on the entire PpCSLD gene family. The loss of function of the CSLD gene family resulted in inhibited protonemal tip growth indicating the CSLD gene family may regulate protonema formation. To further study the function of the CSLD genes in P.patens gene knockout (KO) of the CSLD5 and CSLD8 genes is preformed using CRISPR/Cas9 transformations. Over the summer analysis of the sg1 and sg2 cut sites was preformed on potential CSLD5/8 double knockouts from two different transformations, totaling 140 potential mutants being screened. Screening was preformed using competition-based PCR (cbPCR). When preformed on wild type P.patens the cbPCR primers result in a greater ratio of forward inner primer product (around 564bps) to forward outer primer product (around 840bps), allowing for the screening of mutants by running the cbPCR products on agarose gels and determining the number of base pairs in the final PCR product via electrophoresis with a molecular weight ruler as reference.
Faculty Mentor
Michael Budziszek, Ph.D.
Academic Discipline
College of Arts & Sciences
Repository Citation
Purnell, Henry, "Physcomitrium Patens CSLD Functional Analysis" (2024). Student Research Design & Innovation Symposium. 116.
https://scholarsarchive.jwu.edu/innov_symposium/116