콘텐츠로 건너뛰기
Merck
  • Conditional deletion of SMN in cell culture identifies functional SMN alleles.

Conditional deletion of SMN in cell culture identifies functional SMN alleles.

Human molecular genetics (2020-10-20)
Anton J Blatnik, Vicki L McGovern, Thanh T Le, Chitra C Iyer, Brian K Kaspar, Arthur H M Burghes
초록

Spinal muscular atrophy (SMA) is caused by mutation or deletion of survival motor neuron 1 (SMN1) and retention of SMN2 leading to SMN protein deficiency. We developed an immortalized mouse embryonic fibroblast (iMEF) line in which full-length wild-type Smn (flwt-Smn) can be conditionally deleted using Cre recombinase. iMEFs lacking flwt-Smn are not viable. We tested the SMA patient SMN1 missense mutation alleles A2G, D44V, A111G, E134K and T274I in these cells to determine which human SMN (huSMN) mutant alleles can function in the absence of flwt-Smn. All missense mutant alleles failed to rescue survival in the conditionally deleted iMEFs. Thus, the function lost by these mutations is essential to cell survival. However, co-expression of two different huSMN missense mutants can rescue iMEF survival and small nuclear ribonucleoprotein (snRNP) assembly, demonstrating intragenic complementation of SMN alleles. In addition, we show that a Smn protein lacking exon 2B can rescue iMEF survival and snRNP assembly in the absence of flwt-Smn, indicating exon 2B is not required for the essential function of Smn. For the first time, using this novel cell line, we can assay the function of SMN alleles in the complete absence of flwt-Smn.

MATERIALS
제품 번호
브랜드
제품 설명

Sigma-Aldrich
Anti-α-Tubulin antibody, Mouse monoclonal, clone AA13, purified from hybridoma cell culture
제품번호
팩 크기
재고 정보
가격
수량
Sigma-Aldrich
Anti-SMN2 Antibody, clone SMN-KH, clone SMN-KH, from mouse
제품번호
팩 크기
재고 정보
가격
수량